Prodrugs of aspartyl protease inhibitors

ABSTRACT

The present invention relates to prodrugs of a class of sulfonamides which are aspartyl protease inhibitors. In one embodiment, this invention relates to a novel class of prodrugs of HIV aspartyl protease inhibitors characterized by favorable aqueous solubility, high oral bioavailability and facile in vivo generation of the active ingredient. This invention also relates to pharmaceutical compositions comprising these prodrugs. The prodrugs and pharmaceutical compositions of this invention are particularly well suited for decreasing the pill burden and increasing patient compliance. This invention also relates to methods of treating mammals with these prodrugs and pharmaceutical compositions.

TECHNICAL FIELD OF THE INVENTION

[0001] The present invention relates to prodrugs of a class ofsulfonamides which are aspartyl protease inhibitors. In one embodiment,this invention relates to a novel class of prodrugs of HIV aspartylprotease inhibitors characterized by favorable aqueous solubility, highoral bioavailability and facile in vivo generation of the activeingredient. This invention also relates to pharmaceutical compositionscomprising these prodrugs. The prodrugs and pharmaceutical compositionsof this invention are particularly well suited for decreasing the pillburden and increasing patient compliance. This invention also relates tomethods of treating mammals with these prodrugs and pharmaceuticalcompositions.

BACKGROUND OF THE INVENTION

[0002] Aspartyl protease inhibitors are considered the most effectivecurrent drug in the fight against HIV infection. These inhibitors,however, require certain physicochemical properties in order to achievegood potency against the enzyme. One of these properties is highhydrophobicity. Unfortunately, this property results in poor aqueoussolubility and low oral bioavailability.

[0003] U.S. Pat. No. 5,585,397 describes a class of sulfonamidecompounds that are inhibitors of the aspartyl protease enzyme. Thesecompounds illustrate the drawbacks concomitant to pharmaceuticalcompositions comprising hydrophobic aspartyl protese inhibitors. Forexample, VX-478(4-amino-N-((2-syn,3S)-2-hydroxy-4-phenyl-2((S)-tetrahydrofuran-3-yl-oxycarbonylamino)-butyl-N-isobutyl-benzenesulfonamide)is an aspartyl protease inhibitor disclosed in the '397 patent. In itsmesylate salt form, VX-478 has a relatively low aqueous solubility.While the oral bioavailability of this inhibitor in a “solution”formulation is excellent, the dosage of VX-478 in this form is severelylimited by the amount of liquid present in the particular liquid dosagefrom, e.g., encapsulated into a soft gelatin capsule. A higher aqueoussolubility would increase drug load per unit dosage of VX-478.

[0004] Currently, the mesylate formulation of VX-478 produces an upperlimit of 150 mg of VX-478 in each capsule. Given a therapeutic dose of2400 mg/day of VX-478, this formulation would require a patient toconsume 16 capsules per day. Such a high pill burden would likely resultin poor patient compliance, thus producing sub-optimal therapeuticbenefit of the drug. The high pill burden is also a deterrent toincreasing the amount of the drug administered per day to a patient.Another drawback of the pill burden and the concomitant patientcompliance problem is in the treatment of children infected with HIV.

[0005] Furthermore, these “solution” formulations, such as the mesylatefomulation, are at a saturation solubility of VX-478. This creates thereal potential of having the drug crystallize out of solution undervarious storage and/or shipping conditions. This, in turn, would likelyresult in a loss of some of the oral bioavailability achieved withVX-478.

[0006] One way of overcoming these problems is to develop a standardsolid dosage form, such as a tablet or a capsule or a suspension form.Unfortunately, such solid dosage forms have much lower oralbioavailability of the drug.

[0007] Thus, there is a need to improve the drug load per unit dosageform for aspartyl protease inhibitors. Such an improved dosage formwould reduce the pill burden and increase patient compliance. It wouldalso provide for the possibility of increasing the amounts of the drugadministered per day to a patient.

SUMMARY OF THE INVENTION

[0008] The present invention provides novel prodrugs of a class ofsulfonamide compounds that are inhibitors of aspartyl protease, inparticular, HIV aspartyl protease. These prodrugs are characterized byhigh aqueous solubility, increased bioavailability and are readilymatabolized into the active inhibitors in vivo. The present inventionalso provides pharmaceutical compositions comprising these prodrugs andmethods of treating HIV infection in mammals using these prodrugs andthe pharmaceutical compositions thereof.

[0009] These prodrugs can be used alone or in combination with othertherapeutic or prophylactic agents, such as anti-virals, antibiotics,immunomodulators or vaccines, for the treatment or prophylaxis of viralinfection.

[0010] It is a principal object of this invention to provide novelprodrugs of a class of sulfonamides which are aspartyl proteaseinhibitors, and particularly, HIV aspartyl protease inhibitors. Thisnovel class of sulfonamides is represented by formula I:

[0011] wherein:

[0012] each R¹ is independently selected from the group consisting ofC(O)—, —S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)—and —NR²—C(O)—C(O)—;

[0013] each A is independently selected from the group consisting of 5-7membered monocyclic heterocycles containing from 1-3 endocyclicheteroatoms, which may be optionally methylated at the point ofattachment, optionally benzofused, optionally attached through a C₁-C₃alkyl linker and optionally fused with a 5-7 membered monocyclicheterocycle containing from 1-2 endocyclic heteroatoms, and whereinunmethylated THF is expressly excluded;

[0014] each Ht is independently selected from C₃-C₇ cycloalkyl; C₅-C₇cycloalkenyl; C₆-C₁₀ aryl; or a 5-7 membered saturated or unsaturatedheterocycle, containing one or more heteroatoms selected from N, N(R²),O, S and S(O)_(n); wherein said aryl or said heterocycle is optionallyfused to Q; and wherein any member of said Ht is optionally substitutedwith one or more substituents independently selected from oxo, —OR²,SR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R², —C(O)—N(R²)₂, —S(O)₂—N(R²)₂,—N(R²)—C(O)—R², —C(O)—R², —S(O)_(n)—R², —OCF₃, —S(O)_(n)—Q,methylenedioxy, —N(R²)—S(O)₂(R²), halo, —CF₃, —NO₂, Q, —OQ, —OR⁷, —SR⁷,—R⁷, —N(R²)(R⁷) or —N(R⁷)₂;

[0015] each Q is independently selected from a 3-7 membered saturated,partially saturated or unsaturated carbocyclic ring system; or a 5-7membered saturated, partially saturated or unsaturated heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) orN(R²); wherein Q is optionally substituted with one or more groupsselected from oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)—R², —R²—OH, —CN,—CO₂R², —C(O)—N(R²)₂, halo or —CF₃;

[0016] each R² is independently selected from the group consisting of Hand C₁-C₃ alkyl optionally substituted with Q;

[0017] each x is independently 0 or 1;

[0018] each R³ is independently selected from the group consisting of H,Ht, C₁-C₆ alkyl and C₂-C₆ alkenyl wherein any member of said R³, exceptH, may be optionally substituted with one or more substituents selectedfrom the group consisting of —OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Ht,—CN, —SR², —CO₂R², NR²—C(O)—R²;

[0019] each n is independently 1 or 2;

[0020] G, when present, is selected from H, R⁷ or C₁-C₄ alkyl, or, whenG is C₁-C₄ alkyl, G and R⁷ are bound to one another either directly orthrough a C₁-C₃ linker to form a heterocyclic ring; or

[0021] when G is not present (i.e., when x in (G)_(x) is 0), then thenitrogen to which G is attached is bound directly to the R⁷ group on—OR⁷;

[0022] each D and D′ is independently selected from the group consistingof Q; C₁-C₅ alkyl, which may be optionally substituted with one or moregroups selected from C₃-C₆ cycloalkyl, —OR², —R³, —O—Q, —S—Q and Q;C₂-C₄ alkenyl, which may be optionally substituted with one or moregroups selected from the group consisting of C₃-C₆ cycloalkyl, —OR², R³,O—Q and Q; C₃-C₆ cycloalkyl, which may be optionally substituted with orfused with Q; and C₅-C₆ cycloalkenyl, which may be optionallysubstituted with or fused with R⁶;

[0023] each E is independently selected from the group consisting of Ht;—O—Ht; Ht—Ht; —O—R³; —NR²R³; C₁-C₆ alkyl, which may be optionallysubstituted with one or more groups selected from the group consistingof R⁴ and Ht; and C₂-C₆ alkenyl, which may be optionally substitutedwith one or more groups selected from the group consisting of R⁴ and Ht;C₃-C₆ saturated carbocycle, which is optionally substituted with one ormore groups selected from R⁴ or Ht; or C₅-C₆ unsaturated carbocycle,which is optionally substituted with one or more groups selected from R⁴or Ht;

[0024] each R⁴ is independently selected from the group consisting ofOR², —C(O)—NHR², S(O)₂—NHR², halo, NR²—C(O)—R² and —CN;

[0025] each R⁵ is independently selected from the group consisting of Hand C₁-C₄ alkyl optionally substituted with aryl; and

[0026] each R⁶ is independently selected from the group consisting ofaryl, carbocycle and heterocycle, wherein said aryl, carbocycle orheterocycle may be optionally substituted with one or more groupsselected from the group consisting of oxo, —OR⁵, —R⁵, N(R⁵)(R⁵),N(R⁵)—C(O)—R⁵, —R⁵—OH, —CN, CO₂R⁵, C(O)—N(R⁵)(R⁵), halo and CF₃;

[0027] each R⁷ is independently selected from

[0028] wherein each M is independently selected from H, Li, Na, K, Mg,Ca, Ba, —N(R²)₄, C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, —R⁶; wherein 1 to 4 —CH₂radicals of the alkyl or alkenyl group, other than the —CH₂ that isbound to Z, is optionally replaced by a heteroatom group selected fromO, S, S(O), S(O₂), or N(R²); and wherein any hydrogen in said alkyl,alkenyl or R⁶ is optionally replaced with a substituent selected fromoxo, —OR², —R², N(R²)₂, N(R²)₃, R²OH, —CN, —CO₂R², —C(O)—N(R²)₂,S(O)₂—N(R²)₂, N(R²)—C(O)—R₂, C(O)R², —S(O)_(n)—R², OCF₃, —S(O)_(n)—R⁶,N(R²)—S(O)₂(R²), halo, —CF₃, or —NO₂;

[0029] M′ is H, C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, —R⁶; wherein 1 to 4 —CH₂ ⁻radicals of the alkyl or alkenyl group is optionally replaced by aheteroatom group selected from O, S, S(O), S(O₂), or N(R²); and whereinany hydrogen in said alkyl, alkenyl or R⁶ is optionally replaced with asubstituent selected from oxo, —OR², —R², —N(R²)₂, N(R²)₃, —R²OH, —CN,—CO₂R², —C(O)—N(R²)₂, —S(O)₂—N(R²)₂, —N(R²)—C(O)—R₂, —C(O)R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—R⁶, —N(R²)—S(O)₂(R²), halo, —CF₃, or—NO₂;

[0030] Z is O, S, N(R²)₂, or, when M is absent, H;

[0031] Y is P or S;

[0032] X is O or S; and

[0033] R⁹ is C(R²)₂, O or N(R²); and wherein when Y is S, Z is not S;and

[0034] R⁶ is a 5-6 membered saturated, partially saturated orunsaturated carbocyclic or heterocyclic ring system, or an 8-10 memberedsaturated, partially saturated or unsaturated bicyclic ring system;wherein any of said heterocyclic ring systems contains one or moreheteroatoms selected from O, N, S, S(O)_(n) or N(R²); and wherein any ofsaid ring systems optionally contains 1 to 4 substituents independentlyselected from OH, C₁-C₄ alkyl, O—C₁-C₄ alkyl or OC(O)C₁-C₄ alkyl.

[0035] It is also an object of this invention to provide pharmaceuticalcompositions comprising the sulfonamides of formula I and methods fortheir use as inhibitors of HIV aspartyl protease.

DETAILED DESCRIPTION OF THE INVENTION

[0036] In order that the invention herein described may be more fullyunderstood, the following detailed description is set forth. In thedescription, the following abbreviations are used: Designation Reagentor Fragment Ac acetyl Me methyl Et ethyl Bn benzyl Trityltriphenylmethyl Asn D- or L-asparagine Ile D- or L-isoleucine Phe D- orL-phenylalanine Val D- or L-valine Boc tert-butoxycarbonyl Cbzbenzyloxycarbonyl (carbobenzyloxy) Fmoc 9-fluorenylmethoxycarbonyl DCCdicyclohexylcarbodiimide DIC diisopropylcarbodiimide EDC1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride HOBt1-hydroxybenzotriazole HOSu 1-hydroxysuccinimide TFA trifluoroaceticacid DIEA diisopropylethylamine DBU 1,8-diazabicyclo(5.4.0)undec-7-eneEtOAc ethyl acetate t-Bu tert-butyl iBu iso-butyl DMF dimethylformamideTHP tertrahydropyran THF tetrahydrofuran TMSC1 chlorotrimethylsilaneDMSO dimethylsulfoxide

[0037] The following terms are employed herein:

[0038] Unless expressly stated to the contrary, the terms “SO₂—” and“S(O)₂—” as used herein refer to a sulfone or sulfone derivative (i.e.,both appended groups linked to the S), and not a sulfinate ester.

[0039] The term “backbone” refers to the structural representation of acompound of this invention, as set forth in the figures drawn in thisapplication.

[0040] For the compounds of formula I, and intermediates thereof, thestereochemistry of the —OR⁷ group is defined relative to D on theadjacent carbon atom, when the molecule is drawn in an extended zig-zagrepresentation (such as that drawn for compounds of formula X, XI, XII,XIII, XX, XXI, and XXII). If both —OR⁷ and D reside on the same side ofthe plane defined by the extended backbone of the compound, thestereochemistry of the —OR⁷ bearing carbon atom will be referred to as“syn”. If —OR⁷ and D reside on opposite sides of that plane, thestereochemistry of the —OR⁷ bearing carbon atom will be referred to as“anti”.

[0041] As used herein, the term “alkyl”, alone or in combination withany other term, refers to a straight-chain or branch-chain saturatedaliphatic hydrocarbon radical containing the specified number of carbonatoms, or where no number is specified, preferably from 1-10 and morepreferably from 1-5 carbon atoms. Examples of alkyl radicals include,but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, sec-butyl, tert-butyl, pentyl, isoamyl, n-hexyl and the like.

[0042] The term “alkenyl”, alone or in combination with any other term,refers to a straight-chain or branched-chain mono- or poly-unsaturatedaliphatic hydrocarbon radical containing the specified number of carbonatoms, or where no number is specified, preferably from 2-10 carbonatoms and more preferably, from 2-6 carbon atoms. Examples of alkenylradicals include, but are not limited to, ethenyl, E- and Z-propenyl,isopropenyl, E- and Z-butenyl, E- and Z-isobutenyl, E- and Z-pentenyl,E- and Z-hexenyl, E,E-, E,Z-, Z,E- and Z,Z-hexadienyl and the like.

[0043] The term “aryl”, alone or in combination with any other term,refers to a carbocyclic aromatic radical (such as phenyl or naphthyl)containing the specified number of carbon atoms, preferably from 6-14carbon atoms, and more preferably from 6-10 carbon atoms. Examples ofaryl radicals include, but are not limited to phenyl, naphthyl, indenyl,indanyl, azulenyl, fluorenyl, anthracenyl and the like.

[0044] The term “cycloalkyl”, alone or in combination with any otherterm, refers to a cyclic saturated hydrocarbon radical containing thespecified number of carbon atoms, preferably from 3-7 carbon atoms.Examples of cycloalkyl radicals include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and thelike.

[0045] The term “cycloalkenyl”, alone or in combination with any otherterm, refers to a cyclic hydrocarbon radical containing the specifiednumber of carbon atoms with at least one endocyclic carbon-carbon bond.Where no number of carbon atoms is specified, a cycloalkenyl radicalpreferably has from 5-7 carbon atoms. Examples of cycloalkenyl radicalsinclude, but are not limited to, cyclopentenyl, cyclohexenyl,cyclopentadienyl and the like.

[0046] The term “THF” refers to a tetrahydrofuran ring attached at anyring carbon resulting in a stable structure.

[0047] The term “carbocycle” refers to a stable nonaromatic 3 to8-membered carbon ring which may be saturated, mono-unsaturated orpoly-unsaturated. The carbocycle may be attached at any endocycliccarbon atom which results in a stable structure. Preferred carbocycleshave 5-6 carbons.

[0048] The term “heterocycle”, unless otherwise defined herein, refersto a stable 3-7 membered monocyclic heterocyclic ring or 8-11 memberedbicyclic heterocyclic ring which is either saturated or unsaturated, andwhich may be optionally benzofused if monocyclic. Each heterocycleconsists of one or more carbon atoms and from one to four heteroatomsselected from the group consisting of nitrogen, oxygen and sulfur. Asused herein, the terms “nitrogen and sulfur heteroatoms” include anyoxidized form of nitrogen and sulfur, and the quaternized form of anybasic nitrogen. In addition, any ring nitrogen may be optionallysubstituted with a substituent R², as defined herein for compounds offormula I. A heterocycle may be attached at any endocyclic carbon orheteroatom which results in the creation of a stable structure. Aheterocycle may be attached at any endocyclic carbon or heteroatom whichresults in the creation of a stable structure. Preferred heterocyclesinclude 5-7 membered monocyclic heterocycles and 8-10 membered bicyclicheterocycles. Preferred heterocycles defined above include, for example,benzimidazolyl, imidazolyl, imidazolinoyl, imidazolidinyl, quinolyl,isoquinolyl, indolyl, indazolyl, indazolinolyl, perhydropyridazyl,pyridazyl, pyridyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl,pyrazinyl, quinoxolyl, piperidinyl, pyranyl, pyrazolinyl, piperazinyl,pyrimidinyl, pyridazinyl, morpholinyl, thiamorpholinyl, furyl, thienyl,triazolyl, thiazolyl, β-carbolinyl, tetrazolyl, thiazolidinyl,benzofuranoyl, thiamorpholinyl sulfone, oxazolyl, benzoxazolyl,oxopiperidinyl, oxopyrroldinyl, oxoazepinyl, azepinyl, isoxazolyl,isothiazolyl, furazanyl, tetrahydropyranyl, tetrahydrofuranyl,thiazolyl, thiadiazoyl, dioxolyl, dioxinyl, oxathiolyl, benzodioxolyl,dithiolyl, thiophenyl, tetrahydrothiophenyl and sulfolanyl.

[0049] The term “halo” refers to a radical of fluorine, chlorine,bromine or iodine.

[0050] The term “linker” refers to a structural unit through which twoother moieties are joined. For example, the term “C₁-C₃ alkyl linker”refers to a 1-3 carbon unit which attaches two other moieties together.

[0051] The terms “oxygenated heterocycle” and “heterocycle containingendocyclic oxygen atoms” are used interchangeably and refer to amonocyclic or bicyclic heterocycle containing a specified number ofendocyclic oxygen atoms. Preferably, such oxygenated heterocyclescontain only endocyclic oxygen heteroatoms. Examples of oxygenatedheterocycles, include, but are not limited to: dioxanyl, dioxolanyl,tetrahydrofuranyl, tetrahydrofurodihydrofuranyl, tetrahydropyranyl,tetrahydropyranodihydrofuranyl, dihydropyranyl, tetrahydrofurofuranyland tetrahydropyranofuranyl.

[0052] The terms “HIV protease” and “HIV aspartyl protease” are usedinterchangeably and refer to the aspartyl protease encoded by the humanimmunodeficiency virus type 1 or 2. In a preferred embodiment of thisinvention, these terms refer to the human immunodeficiency virus type 1aspartyl protease.

[0053] The term “antiviral agent” or “anti-retroviral agent” refers to acompound or drug which possesses viral inhibitory activity. Such agentsinclude reverse transcriptase inhibitors (including nucleoside andnon-nucleoside analogs) and protease inhibitors. Preferably the proteaseinhibitor is an HIV protease inhibitor. Examples of nucleoside analogreverse transcriptase inhibitors include, but are not limited to,zidovudine (AZT), dideoxycytidine (ddC), didanosine (ddI), stavudine(d4T), 3TC, 935U83, 1592U89 and 524W91. Examples of non-nucleosideanalog reverse transcriptase inhibitors include, but are not limited todelavirdine (U90) and nevirapine. Examples of HIV protease inhibitorsinclude, but are not limited to, saquinavir (Ro 318959), L-735,524, ABT538 (A80538), AG 1343, XM 412, XM 450, BMS 186318 and CPG 53,437.

[0054] The term “pharmaceutically effective amount” refers to an amounteffective in treating HIV infection in a patient either as monotherapyor in combination with other agents. The term “treating” as used hereinrefers to the alleviation of symptoms of a particular disorder in apatient or the improvement of an ascertainable measurement associatedwith a particular disorder. Specifically, with respect to HIV, effectivetreatment using the compounds and compositions of this invention wouldresult in an improvement in an HIV associated ascertainable measurement.Such measurements include, but are not limited to, reduction in viralload in plasma or another defined tissue compartment as measured by,e.g. RT-PCR or branched-chain DNA PCR or culturable virus measurements,□-2 microglobulin or p24 levels, number of CD₄ ⁺ cells or ratio of CD₄⁺/CD₈ ⁺ cells, or functional markers such as improvement in quality oflife or ability to carry out normal functions or reduction inimmunosuppression-related effects. The term “prophylactically effectiveamount” refers to an amount effective in preventing HIV infection in apatient. As used herein, the term “patient” refers to a mammal,including a human.

[0055] The term “pharmaceutically acceptable carrier or adjuvant” refersto a carrier or adjuvant that may be administered to a patient, togetherwith a compound of this invention, and which does not destroy thepharmacological activity thereof and is nontoxic when administered indoses sufficient to deliver a therapeutic amount of the antiretroviralagent.

[0056] The term “point of attachment” refers to the atom through which amoiety is attached to a specified structure. When a point of attachmentmay be optionally methylated, the point of attachment is the carbon atomthrough which a moiety is attached to a specified structure.

[0057] The term “substituted”, whether express or implied and whetherpreceded by the term “optionally” or not, refers to the replacement ofone or more hydrogen radicals in a given structure with the radical of aspecified substituent. When more than one position in a given structuremay be substituted with a substituent selected from a specified group,the substituents may be either the same or different at every position.Typically, when a structure may be optionally substituted, 0-3substitutions are preferred, and 0-1 substitution is most preferred.Most preferred substituents are those which enhance protease inhibitoryactivity or intracellular antiviral activity in permissive mammaliancells or immortalized mammalian cell lines, or which enhancedeliverability by enhancing solubility characteristics or enhancingpharmacokinetic or pharmacodynamic profiles as compared to theunsubstituted compound. Other most preferred substituents include thoseused in the compounds shown in Table I.

[0058] Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from pharmaceutically acceptableinorganic and organic acids and bases. Examples of suitable acidsinclude hydrochloric, hydrobromic, sulfuric, nitric, perchloric,fumaric, maleic, phosphoric, glycollic, lactic, salicylic, succinic,p-toluenesulfonic, tartaric, acetic, citric, methanesulfonic,ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic andbenzenesulfonic acids. Preferred acids include hydrochloric, sulfuric,methanesulfonic and ethanesulfonic acids. Methanesulfonic acid is mostpreferred. Other acids, such as oxalic, while not in themselvespharmaceutically acceptable, may be employed in the preparation of saltsuseful as intermediates in obtaining the compounds of the invention andtheir pharmaceutically acceptable acid addition salts.

[0059] Salts derived from appropriate bases include alkali metal (e.g.,sodium), alkaline earth metal (e.g., magnesium), ammonium and N—(C₁₋₄alkyl)₄ ⁺ salts.

[0060] The term “thiocarbamates” refers to compounds containing thefunctional group N—SO₂—O.

[0061] The compounds of this invention contain one or more asymmetriccarbon atoms and thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers. Allsuch isomeric forms of these compounds are expressly included in thepresent invention. Each stereogenic carbon may be of the R or Sconfiguration. The explicitly shown hydroxyl is also preferred to be synto D, in the extended zig-zag conformation between the nitrogens shownin compounds of formula I.

[0062] Combinations of substituents and variables envisioned by thisinvention are only those that result in the formation of stablecompounds. The term “stable”, as used herein, refers to compounds whichpossess stability sufficient to allow manufacture and which maintainsthe integrity of the compound for a sufficient period of time to beuseful for the purposes detailed herein (e.g., therapeutic orprophylactic administration to a mammal or for use in affinitychromatography applications). Typically, such compounds are stable at atemperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

[0063] The compounds of the present invention may be used in the form ofsalts derived from inorganic or organic acids. Included among such acidsalts, for example, are the following: acetate, adipate, alginate,aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate,camphorate, camphorsulfonate, cyclopentanepropionate, digluconate,dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate,glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate,pectinate, persulfate, 3-phenylpropionate, picrate, pivalate,propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate.

[0064] This invention also envisions the quaternization of any basicnitrogen-containing groups of the compounds disclosed herein. The basicnitrogen can be quaternized with any agents known to those of ordinaryskill in the art including, for example, lower alkyl halides, such asmethyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkylsulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; longchain halides such as decyl, lauryl, myrist_ and stearyl chlorides,bromides and iodides; and aralkyl halides including benzyl and phenethylbromides. Water or oil-soluble or dispersible products may be obtainedby such quaternization.

[0065] The novel sulfonamides of this invention are those of formula I:

[0066] wherein:

[0067] each R¹ is independently selected from the group consisting ofC(O)—, —S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)—and —NR²—C(O)—C(O)—;

[0068] each A is independently selected from the group consisting of 5-7membered monocyclic heterocycles containing from 1-3 endocyclicheteroatoms, which may be optionally methylated at the point ofattachment, optionally benzofused, optionally attached through a C₁-C₃alkyl linker and optionally fused with a 5-7 membered monocyclicheterocycle containing from 1-2 endocyclic heteroatoms, and whereinunmethylated THF is expressly excluded;

[0069] each Ht is independently selected from C₃-C₇ cycloalkyl; C₅-C₇cycloalkenyl; C₆-C₁₀ aryl; or a 5-7 membered saturated or unsaturatedheterocycle, containing one or more heteroatoms selected from N, N(R²),O, S and S(O)_(n); wherein said aryl or said heterocycle is optionallyfused to Q; and wherein any member of said Ht is optionally substitutedwith one or more substituents independently selected from oxo, —OR²,SR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R², —C(O)—N(R²)₂, —S(O)₂—N(R²)₂,—N(R²)—C(O)—R², —C(O)—R², —S(O)_(n)—R², —OCF₃, —S(O)_(n)—Q,methylenedioxy, —N(R²)—S(O)₂(R²), halo, —CF₃, —NO₂, Q, —OQ, —OR⁷, —SR⁷,—R⁷, —N(R²)(R⁷) or —N(R⁷)₂;

[0070] each Q is independently selected from a 3-7 membered saturated,partially saturated or unsaturated carbocyclic ring system; or a 5-7membered saturated, partially saturated or unsaturated heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) orN(R²); wherein Q is optionally substituted with one or more groupsselected from oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)—R², —R²—OH, —CN,—CO₂R², —C(O)—N(R²)₂, halo or —CF₃;

[0071] each R² is independently selected from the group consisting of Hand C₁-C₃ alkyl optionally substituted with Q;

[0072] each x is independently 0 or 1;

[0073] each R³ is independently selected from the group consisting of H,Ht, C₁-C₆ alkyl and C₂-C₆ alkenyl wherein any member of said R³, exceptH, may be optionally substituted with one or more substituents selectedfrom the group consisting of —OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Ht,—CN, —SR², —CO₂R², NR²—C(O)—R²;

[0074] each n is independently 1 or 2;

[0075] G, when present, is selected from H, R⁷ or C₁-C₄ alkyl, or, whenG is C₁-C₄ alkyl, G and R⁷ are bound to one another either directly orthrough a C₁-C₃ linker to form a heterocyclic ring; or

[0076] when G is not present (i.e., when x in (G)_(x) is 0), then thenitrogen to which G is attached is bound directly to the R⁷ group on—OR⁷;

[0077] each D and D′ is independently selected from the group consistingof Q; C₁-C₅ alkyl, which may be optionally substituted with one or moregroups selected from C₃-C₆ cycloalkyl, —OR², —R³, —O—Q, —S—Q and Q;C₂-C₄ alkenyl, which may be optionally substituted with one or moregroups selected from the group consisting of C₃-C₆ cycloalkyl, —OR², R³,O—Q and Q; C₃-C₆ cycloalkyl, which may be optionally substituted with orfused with Q; and C₅-C₆ cycloalkenyl, which may be optionallysubstituted with or fused with R⁶;

[0078] each E is independently selected from the group consisting of Ht;—O—Ht; Ht—Ht; —O—R³; —NR²R³; C₁-C₆ alkyl, which may be optionallysubstituted with one or more groups selected from the group consistingof R⁴ and Ht; and C₂-C₆ alkenyl, which may be optionally substitutedwith one or more groups selected from the group consisting of R⁴ and Ht;C₃-C₆ saturated carbocycle, which is optionally substituted with one ormore groups selected from R⁴ or Ht; or C₅-C₆ unsaturated carbocycle,which is optionally substituted with one or more groups selected from R⁴or Ht;

[0079] each R⁴ is independently selected from the group consisting ofOR², —C(O)—NHR², S(O)₂—NHR², halo, NR²—C(O)—R² and —CN;

[0080] each R⁵ is independently selected from the group consisting of Hand C₁-C₄ alkyl optionally substituted with aryl; and

[0081] each R⁶ is independently selected from the group consisting ofaryl, carbocycle and heterocycle, wherein said aryl, carbocycle orheterocycle may be optionally substituted with one or more groupsselected from the group consisting of oxo, —OR⁵, —R⁵, N(R⁵)(R⁵),N(R⁵)—C(O)—R⁵, —R⁵—OH, —CN, CO₂R⁵, C(O)—N(R⁵)(R⁵), halo and CF₃;

[0082] each R is independently selected from

[0083] wherein each M is independently selected from H, Li, Na, K, Mg,Ca, Ba, —N(R²)₄, C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, —R⁶; wherein 1 to 4 —CH₂radicals of the alkyl or alkenyl group, other than the —CH₂ that isbound to Z, is optionally replaced by a heteroatom group selected fromO, S, S(O), S(O₂), or N(R²); and wherein any hydrogen in said alkyl,alkenyl or R⁶ is optionally replaced with a substituent selected fromoxo, —OR², —R², N(R²)₂, N(R²)₃, R²OH, —CN, —CO₂R², —C(O)—N(R²)₂,S(O)₂—N(R²)₂, N(R²)—C(O)—R₂, C(O)R², —S(O)_(n)—R², OCF₃, —S(O)_(n)—R⁶,N(R²)—S(O)₂(R²), halo, —CF₃, or —NO₂;

[0084] M′ is H, C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, —R⁶; wherein 1 to 4 —CH₂radicals of the alkyl or alkenyl group is optionally replaced by aheteroatom group selected from O, S, S(O), S(O₂), or N(R²); and whereinany hydrogen in said alkyl, alkenyl or R⁶ is optionally replaced with asubstituent selected from oxo, —OR², —R², —N(R²)₂, N(R²)₃, —R²OH, —CN,—CO₂R², —C(O)—N(R²)₂, —S(O)₂—N(R²)₂, —N(R²)—C(O)—R₂, —C(O)R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—R⁶, —N(R²)—S(O)₂(R²), halo, —CF₃, or—NO₂;

[0085] Z is O, S, N(R²)₂, or, when M is absent, H;

[0086] Y is P or S;

[0087] X is O or S; and

[0088] R⁹ is C(R²)₂, O or N(R²); and wherein when Y is S, Z is not S;and

[0089] R⁶ is a 5-6 membered saturated, partially saturated orunsaturated carbocyclic or heterocyclic ring system, or an 8-10 memberedsaturated, partially saturated or unsaturated bicyclic ring system;wherein any of said heterocyclic ring systems contains one or moreheteroatoms selected from O, N, S, S(O)_(n) or N(R²); and wherein any ofsaid ring systems optionally contains 1 to 4 substituents independentlyselected from OH, C₁-C₄ alkyl, O—C₁-C₄ alkyl or OC(O)C₁-C₄ alkyl.

[0090] Preferred compounds of formula I have the following definitionsfor one or more of the below-specified substituents:

[0091] each R¹ is —O—C—(O)—;

[0092] each A is independently selected from the group consisting of 5-6membered monocyclic heterocycles containing from 1-2 endocyclic oxygenatoms, which may be optionally methylated at the point of attachment,optionally attached through a C₁-C₃ alkyl linker and optionally fusedwith a 5-6 membered monocyclic heterocycle containing from 1-2endocyclic oxygen atoms, and more preferably, A is selected from thegroup consisting of dioxanyl (preferably, 1,3-dioxanyl), dioxolanyl,dioxolanylmethyl, 3-methyl THF, tetrahydrofurofuranyl,tetrahydrofurodihydrofuranyl, tetrahydropyranofuranyl,tetrahydropyranodihydrofuranyl, pyranyl, dihydropyranyl andtetrahydropyranyl. Most preferably, A is 1,3-dioxanyl attached at the5-position.

[0093] each D is C₁-C₅ alkyl, which may be optionally substituted withone or more Ht, more preferably D is C₁-C₅ alkyl, which may beoptionally substituted with one group selected from C₆-C₁₀ aryl andC₃-C₆ cycloalkyl, even more preferably D is selected from benzyl,isobutyl, cyclopentylmethyl, and cyclohexylmethyl and most preferably, Dis benzyl or isobutyl;

[0094] each D′ is selected from the group consisting of C₁-C₆ alkyloptionally substituted with R⁶ (wherein each R⁶ is independentlyselected from the group consisting of carbocycle and heterocycle,wherein said heterocycle or carbocycle may be optionally substitutedwith one or more groups selected from the group consisting of oxo, OR⁵,—R⁵, N(R⁵)(R⁵), N(R⁵)—C(O)—R⁵, —R⁵—OH, —CN, CO₂R⁵, C(O)—N(R⁵)(R⁵), haloand CF₃ and each R⁵ is independently selected from the group consistingof H and C₁-C₃ alkyl), and more preferably D′ is selected from the groupconsisting of C₁C₄ alkyl optionally substituted with one 3-6 memberedcarbocycle or one 5-6 membered heterocycle, and most preferably, D′ isselected from the group consisting of isobutyl, cyclopentylmethyl andcyclohexylmethyl;

[0095] each E is Ht and more preferably, E is phenyl substituted with0-2 substituents chosen from the group consisting of OH, OR⁷ OCH₃, NH₂,NHCOCH₃, SCH₃, and CH₃; or phenyl fused with 5-6 membered heterocycle,and even more preferably, E is phenyl substituted with one substituentselected from the group consisting of OH, OR⁷ OCH₃, NH₂, NHCOCH₃, SCH₃,and CH₃; or phenyl fused with 5-6 membered heterocycle, and mostpreferably, E is phenyl substituted with NH₂, NHR⁷ or N(R⁷)₂ (preferablyin the meta- or para-position).

[0096] Preferably R⁷ is

[0097] PO₃K₂, PO₃Ca, PO₃-spermine, P₃-(spermidine)₂ or PO₃-(meglamine)₂.

[0098] It will be understood by those of skill in the art that componentM or M′ in the formulae set forth herein will have either a covalent, acovalent/zwitterionic, or an ionic association with either Z or R⁹depending upon the actual choice for M or M′. When M or M′ is hydrogen,alkyl, alkenyl, or R⁶, M or M′ is covalently bound to R⁹ or Z. If M is amono- or bivalent metal or other charged species (i.e., NH₄ ⁺), there isan ionic interaction between M and Z and the resulting compound is asalt.

[0099] When x is 0 in (M)_(x), Z may be a charged species. When thatoccurs, the other M may be oppositely charged to produce a 0 net chargeon the molecule. Alternatively, the counter ion may located elsewhere inthe molecule.

[0100] Except where expressly provided to the contrary, as used herein,the definitions of variables A, R¹-R⁴, R⁶-R⁹, Ht, B, x, n, D, D′, M, Q,X, Y, Z and E are to be taken as they are defined above for thecompounds of formula I.

[0101] Table I illustrates preferred compounds of this invention: TABLE1

COMPOUND A D D′ E 1

2

3

4

5

6

7 (Isomer A)

8 (Isomer B)

9 (Isomer A)

10 (Isomer B)

11

—CH₃

12

13

14

15

16

[0102] The prodrugs of the present invention may be synthesized usingconventional synthetic techniques. WO 96/33187 discloses the synthesisof compounds of formula:

[0103] wherein A, R¹, D, D′ and E are as defined above. Prodrugs offormula (I) of the present invention can be readily synthesized from the'187 compounds using conventional techniques. One of skill in the artwould be well aware of conventional synthetic reagents to convert the—OH group of the '187 compounds to a desired —OR⁷ functionality of thepresent invention, wherein R⁷ is as defined above. The relative easewith which the compounds of this invention can be synthesized representsan enormous advantage in the large scale production of these compounds.

[0104] For example, VX-478, a compound disclosed in U.S. Pat. No.5,585,397, can be readily converted to the corresponding bis-phosphateester derivative, as shown below:

[0105] Alternatively, if the monophosphate ester of VX-478 is desired,then the synthetic scheme can be readily adapted by beginning with the4-nitrophenyl derivative of VX-478, as shown below:

[0106] Although unmethylated tetrahydrofuran embodiments of formula I,such as VX 478, are expressly excluded from the present invention, oneof skill in the art would readily be able to prepare the correspondingmonophosphate and bis-phosphate esters of the present invention usingsimilar reaction conditions.

[0107] Further examples of specific compounds which may be converted tothe prodrugs of this invention by similar techniques (and the synthesesof those intermediates to the compounds of the present invention) aredisclosed in WO 94/05639 and '397 patent, the disclosures of which areherein incorporated by reference.

[0108] Pharmaceutically acceptable salts of the compounds of the presentinvention may be readily prepared using known techniques. For example,the disodium salt of the mono-phosphate ester shown above can beprepared as shown below:

[0109] The compounds of this invention may be modified by appendingappropriate functionalities to enhance selective biological properties.Such modifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

[0110] Without being bound by theory, we believe that two differentmechanisms are involved in converting the prodrugs of this inventioninto the active drug, depending upon the structure of the prodrug. Thefirst mechanism involves the enzymatic or chemical transformation of theprodrug species into the active form. The second mechanism involves theenzymatic or chemical cleavage of a functionality on the prodrug toproduce the active compound.

[0111] The chemical or enzymatic transformation can involve to transferof a functional group (i.e., R⁷) from one heteroatom within the moleculeto another heteroatom. This transfer is demonstrated in the chemicalreactions shown below:

[0112] The cleavage mechanism is demonstrated by the reaction belowwhere a phosphate ester-containing prodrug is converted into the activeform of the drug by removal of the phosphate group.

[0113] These protease inhibitors and their utility as inhibitors ofaspartyl proteases are described in WO 96/33187, the disclosure of whichis incorporated herein by reference.

[0114] The prodrugs of the present invention are characterized byunexpectedly high aqueous solubility. This solubility facilitatesadministration of higher doses of the prodrug, resulting in a greaterdrug load per unit dosage. The prodrugs of the present invention arealso characterized by facile hydrolytic cleavage to release the activeaspartyl protease inhibitor in vivo. The high aqueous solubility and thefacile in vivo metabolism result in a greater bioavailability of thedrug. As a result, the pill burden on a patient is significantlyreduced.

[0115] The prodrugs of this invention may be employed in a conventionalmanner for the treatment of viruses, such as HIV and HTLV, which dependon aspartyl proteases for obligatory events in their life cycle. Suchmethods of treatment, their dosage levels and requirements may beselected by those of ordinary skill in the art from available methodsand techniques. For example, a prodrug of this invention may be combinedwith a pharmaceutically acceptable adjuvant for administration to avirally-infected patient in a pharmaceutically acceptable manner and inan amount effective to lessen the severity of the viral infection.

[0116] Alternatively, the prodrugs of this invention may be used invaccines and methods for protecting individuals against viral infectionover an extended period of time. The prodrugs may be employed in suchvaccines either alone or together with other compounds of this inventionin a manner consistent with the conventional utilization of proteaseinhibitors in vaccines. For example, a prodrug of this invention may becombined with pharmaceutically acceptable adjuvants conventionallyemployed in vaccines and administered in prophylactically effectiveamounts to protect individuals over an extended period time against HIVinfection. As such, the novel protease inhibitors of this invention canbe administered as agents for treating or preventing HIV infection in amammal.

[0117] The prodrugs of this invention may be administered to a healthyor HIV-infected patient either as a single agent or in combination withother anti-viral agents which interfere with the replication cycle ofHIV. By administering the compounds of this invention with otheranti-viral agents which target different events in the viral life cycle,the therapeutic effect of these compounds is potentiated. For instance,the co-administered anti-viral agent can be one which targets earlyevents in the life cycle of the virus, such as cell entry, reversetranscription and viral DNA integration into cellular DNA. Anti-HIVagents targeting such early life cycle events include, didanosine (ddI),alcitabine (ddC), d4T, zidovudine (AZT), polysulfated polysaccharides,sT4 (soluble CD4), ganiclovir, dideoxycytidine, trisodiumphosphonoformate, eflornithine, ribavirin, acyclovir, alpha interferonand trimenotrexate. Additionally, non-nucleoside inhibitors of reversetranscriptase, such as TIBO or nevirapine, may be used to potentiate theeffect of the compounds of this invention, as may viral uncoatinginhibitors, inhibitors of trans-activating proteins such as tat or rev,or inhibitors of the viral integrase.

[0118] Combination therapies according to this invention exert asynergistic effect in inhibiting HIV replication because each componentagent of the combination acts on a different site of HIV replication.The use of such combinations also advantageously reduces the dosage of agiven conventional anti-retroviral agent which would be required for adesired therapeutic or prophylactic effect as compared to when thatagent is administered as a monotherapy. These combinations may reduce oreliminate the side effects of conventional single anti-retroviral agenttherapies while not interfering with the anti-retroviral activity ofthose agents. These combinations reduce potential of resistance tosingle agent therapies, while minimizing any associated toxicity. Thesecombinations may also increase the efficacy of the conventional agentwithout increasing the associated toxicity. In particular, we havediscovered that these prodrugs act synergistically in preventing thereplication of HIV in human T cells. Preferred combination therapiesinclude the administration of a prodrug of this invention with AZT, ddI,ddC or d4T.

[0119] Alternatively, the prodrugs of this invention may also beco-administered with other HIV protease inhibitors such as Ro 31-8959(Roche), L-735,524 (Merck), XM 323 (Du-Pont Merck) and A-80,987 (Abbott)to increase the effect of therapy or prophylaxis against various viralmutants or members of other HIV quasi species.

[0120] We prefer administering the prodrugs of this invention as singleagents or in combination with retroviral reverse transcriptaseinhibitors, such as derivatives of AZT, or other HIV aspartyl proteaseinhibitors. We believe that the co-administration of the compounds ofthis invention with retroviral reverse transcriptase inhibitors or HIVaspartyl protease inhibitors may exert a substantial synergistic effect,thereby preventing, substantially reducing, or completely eliminatingviral infectivity and its associated symptoms.

[0121] The prodrugs of this invention can also be administered incombination with immunomodulators (e.g., bropirimine, anti-human alphainterferon antibody, IL-2, GM-CSF, methionine enkephalin, interferonalpha, diethyldithiocarbamate, tumor necrosis factor, naltrexone andrEPO); and antibiotics (e.g., pentamidine isethiorate) to prevent orcombat infection and disease associated with HIV infections, such asAIDS and ARC.

[0122] When the prodrugs of this invention are administered incombination therapies with other agents, they may be administeredsequentially or concurrently to the patient. Alternatively,pharmaceutical or prophylactic compositions according to this inventionmay be comprised of a combination of a prodrug of this invention andanother therapeutic or prophylactic agent.

[0123] Although this invention focuses on the use of the prodrugsdisclosed herein for preventing and treating HIV infection, thecompounds of this invention can also be used as inhibitory agents forother viruses which depend on similar aspartyl proteases for obligatoryevents in their life cycle. These viruses include, as well as otherAIDS-like diseases caused by retroviruses, such as simianimmunodeficiency viruses, but are not limited to, HTLV-I and HTLV-II. Inaddition, the compounds of this invention may also be used to inhibitother aspartyl proteases, and in particular, other human aspartylproteases, including renin and aspartyl proteases that processendothelin precursors.

[0124] Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle. Pharmaceutically acceptable carriers, adjuvants and vehiclesthat may be used in the pharmaceutical compositions of this inventioninclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

[0125] The pharmaceutical compositions of this invention may beadministered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir. Weprefer oral administration or administration by injection. Thepharmaceutical compositions of this invention may contain anyconventional non-toxic pharmaceutically-acceptable carriers, adjuvantsor vehicles. The term parenteral as used herein includes subcutaneous,intracutaneous, intravenous, intramuscular, intra-articular,intrasynovial, intrasternal, intrathecal, intralesional and intracranialinjection or infusion techniques.

[0126] The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant such as Ph. Helv or a similar alcohol.

[0127] The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carriers which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried corn starch. Whenaqueous suspensions are administered orally, the active ingredient iscombined with emulsifying and suspending agents. If desired, certainsweetening and/or flavoring and/or coloring agents may be added.

[0128] The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

[0129] Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible by topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esterswax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable enema formulation. Topically-transdermal patches arealso included in this invention.

[0130] The pharmaceutical compositions of this invention may beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other solubilizing or dispersingagents known in the art.

[0131] Dosage levels of between about 0.01 and about 100 mg/kg bodyweight per day, preferably between about 0.5 and about 50 mg/kg bodyweight per day of the active ingredient compound are useful in theprevention and treatment of viral infection, including HIV infection.Typically, the pharmaceutical compositions of this invention will beadministered from about 1 to about 5 times per day or alternatively, asa continuous infusion. Such administration can be used as a chronic oracute therapy. The amount of active ingredient that may be combined withthe carrier materials to produce a single dosage form will varydepending upon the host treated and the particular mode ofadministration. A typical preparation will contain from about 5% toabout 95% active compound (w/w), Preferably, such preparations containfrom about 20% to about 80% active compound.

[0132] Upon improvement of a patient's condition, a maintenance dose ofa compound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease.Patients may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

[0133] As the skilled artisan will appreciate, lower or higher dosesthan those recited above may be required. Specific dosage and treatmentregimens for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health status, sex, diet, time ofadministration, rate of excretion, drug combination, the severity andcourse of the infection, the patient's disposition to the infection andthe judgment of the treating physician.

[0134] In order that this invention be more fully understood, thefollowing examples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

EXAMPLE 1

[0135] General Conditions:

[0136] (A) Analytical HPLC 0-100%B/30 min, 1.5 mL/min, A=0.1% TFA inwater, B=0.1% TFA in acetonitrile. Detection at 254 and 220 nm, C18reverse phase Vydac, t0=2.4 min.

[0137] (B) ⅓ v/v EtOAc/hexane

[0138] (C) ½ v/v EtOAc/hexane

[0139] (D) Analytical HPLC 0-100%B/10 min, 1.5 mL/min, A=0.1% TFA inwater, B=0.1% TFA in acetonitrile. Detection at 254 and 220 nm, C18reverse phase Vydac, t₀=2.4 min.

[0140] A mixture of 2.0 g (3.7 mMol) of 197 and 3.0 g (16 mMol) ofdi-p-nitrophenyl carbonate in 10 ml of dimethylformamide was treated at25° with 4 ml (4 mMol) of P4-phosphazene base (Fluka, 1M in hexane). Themixture was stirred for 6 h at 25° until all of the starting alcohol wasconsumed. The reaction mixture was partitioned between ethyl acetate and1N hydrochloric acid. The organic layer was washed with 1N sodiumhydroxide and brine, dried over magnesium sulfate and concentrated invacuo. Titration with dichloromethane gave the desired mixed carbonate(1.2 g crop1 and 0.6 g crop 2) as a fine powder. Combined yield: 69%.Rf=0.13 (⅓ EtOAc/hexane, conditions B), Rf=0.40 (½ EtOAc/hexane,conditions C), tHPLC=23.83 min (A), MS(ES+) 701 (M+1).

[0141] 1H-NMR (CDCl3): 0.82 (6H,dd), 1.9 (2H,m), 2.15 (1H,m), 2.8(1H,m), 3.0 (4H,m), 3.5 (2H,m), 3.6 (1H,m), 3.8 (4H,m), 4.3 (1H,bs), 4.8(1H,m), 5.17 (2H,m), 7.7 (7H,m), 7.95 (2H,d), 8.35 (4H,m).

[0142] 13C (CDCl3): 155.2 152.2, 149.9, 145.6, 135.9, +129.0, +128.8,+128.5, +127.2, +125.4, +124.4, +121.8, +78.1, +75.8, −73.1, −66.9,−56.5, +52.7, −48.2, −35.9, −35.9, 32.6, −+26.4, +19.9, +19.8.

EXAMPLE 2

[0143]

[0144] To 0.20 g (0.286 mM) of 198 dissolved in 3 ml of THF was added0.11 g (1.14 mM) of 1-Methyl-piperidine and the mixture was stirredovernight at room temperature (“rt”). All the solvents were thenevaporated and the solid residue partitioned between EtOAc and water.The volatiles were removed and, where appropriate, the residue wastreated with 1:1 TFA/DCM over 30 min at rt to remove the Boc protectinggroup. The product was dissolved in 0.25 ml TFA and 1.5 ml THF.Hydrogenolysis for 10 hours in presence of 30 mg of 10% Pd/C gave thedesired compound. The final purification was on preparative reversedphase C18 using conditions Example 1, except that the flow rate was 18ml/min.

[0145] C,H,N: calc: 49.27, 5.57, 8.25, found 49.15, 5.76, 8.29C₃₁H₄₅N₅O₇S₁. 1.9CF₃COOH

[0146] LC/MS (ES+) 632 (M+1) 1 peak at 4.71 min

[0147] Analytical HPLC(A) t=N/A min

[0148] 1H:0.71 (3H,d), 0.74 (3H,d), 1.80 (2H,m), 2.03 (1H,m), 2.63(2H,m), 2.74 (1H,m), 2.82 (3H,s), 2.92 (2H,m), 3.20 (4H,m), 3.42 (3H,m),3.62 (2H,m), 3.75 (1H,m), 4.05 (3H,m), 4.97 (2H,m), 6.2 (1H,bs), 6.60(2H,m), 7.22 (5H,m), 7.40 (3H,m), 13C (DMSO): 156.4, 154.0, 153.8,138.8, 129.6, 129.5, 128.3, 126.5, 123.7, 112.7, 74.8, 72.9, 66.7, 58.2,54.0, 53.1, 49.3, 42.3, 40.8, 36.0, 33.3, 25.8, 20.4, 20.3

EXAMPLE 3

[0149]

[0150] The synthesis of compound 200 from compound 198 was carried asdescribed in Example 1, except that N,N-dimethyl-aminoethanol was usedin place of di-p-nitrophenyl carbonate.

[0151] 1HNMR (acetone-d6): 0.82 (6H,dd), 1.83 (2H,m), 2.07 (1H,m), 2.64(2H,m), 2.82 (6H,s), 2.90 (2H,m), 3.19 (1H,m), 3.38 (4H,m), 3.63 (2H,m),3.76 (1H,m), 4.17 (2YH,m), 4.40 (1H,m), 4.56 (1H,m), 4.96 (1H,m), 5.06(1H,m), 6.06 (1H,d), 6.68 (2H,d), 7.23 (5H,m), 7.47 (2H,d).

[0152] 13CNMR (acetone d6): 20.2, 20.3, 27.5, 33.4, 35.6, 43.8, 50.1,54.2, 56.4, 58.5, 63.1, 67.4, 73.6, 76.2, 79.9, 114.2, 118.3, 127.4,129,2, 130.1, 130.3, 139.3, 153.4, 157.0.

[0153] LC/MS: 1 peak, 621 (MH+).

EXAMPLE 4

[0154]

[0155] The synthesis of compound 201 from compound 198 was carried asdescribed in Example 1, except that N-acetyl-ethylenediamine was used inplace of di-p-nitrophenyl carbonate,

[0156] C,H,N: calc: 49.66, 5.64, 8.83, found 49.76, 5.98, 8.93C₃₀H₄₃N₅O₈S₁. 1.4CF₃COOH.

[0157] LC/MS (ES+) 634 (M+1) 1 peak at 5.08 min.

[0158] Analytical HPLC(A)⁻ t=15.92 min.

[0159] 1H: d-3 acetonitrile: 0.88 (6H,dd), 1.92 (3H,s), 1.94 (2H,m),2.17 (1H,m), 2.72 (2H,m), 2.96 (2H,m), 3.07 (3H,m), 3.29 (1H,m), 3.42(3H,m), 3.69 (1H,m), 3.77 (1H,m), 3.82 (1H,m), 4.133 (1H,m), 4.40(1H,bs), 5.05 (2H,m), 5.80 (1H,m), 6.10 (1H,d), 6.78 (2H,d), 6.83(1H,bs), 7.28 (5H,m), 7.58 (2H,d).

[0160] 13C (d3-acetonitrile): 157.1, 157.0, 153.2, 139.6, +130.3,+130.2, +129.2, +127.2, 126.2, +114.2, +76.0, +75.4, −73.6, −67.4,−58.2, +54.9, −50.2, −41.6, −39.8, −35.9, −33.4, +27.3, +23.1, +20.4,+20.2.

EXAMPLE 5

[0161]

[0162] The synthesis of compound 202 from compound 198 was carried asdescribed in Example 1, except that mono N-Boc-piperazine was used inplace of di-p-nitrophenyl carbonate.

[0163] C,H,N: calc: 48.28, 5.68, 8.41, found 48.28, 5.36, 8.28C₃₀H₄₃N₅O₇S₁×2 CF₃COOH

[0164] LC/MS (ES+) 618 (M+1) 1 peak at 4.36 min.

[0165] Analytical HPLC(A) t=14.84 min.

[0166] 1H: d6-DMSO: 0.72 (3H,d), 0.77 (3H,d), 1.78 (2H,m), 2.09 (1H,m),2.64 (2H,m), 2.73 (1H,m), 2.80 (1H,m), 3.08 (4H,m), 3.32 (2H,m), 3.41(1H,m), 3.50 (4H,m), 3.54 (1H,m), 3.63 (1H,m), 3.70 (1H,m), 3.98 (1H,m),4.89 (1H,m), 4.97 (1H,m), 6.61 (2H,d), 7.23 (5H,m), 7.42 (3H,m), 8.88(2H,bs).

[0167] 13C: (DMSO): 155.7, 153.6, 153.0, 138.4, +129.1, +129.0, +128.1,+126.1, 123.2, +112.7, +75.2, +74.4, −72.5, −66.2, −56.9, +53.1, −48.8,−42.5, −40.8, −35.0, −32.2, +26.2, +20.0, +19.8.

EXAMPLE 6

[0168]

[0169] The synthesis of compound 203 from compound 198 was carried asdescribed in Example 1, except that mono-N-Boc-ethylenediamine was usedin place of di-p-nitrophenyl carbonate.

[0170] C,H,N: calc: 46.89, 5.29, 8.54, found 46.50, 5.51, 8.54C₂₈H₄₁N₅O₇S₁×2 CF₃COOH.

[0171] LC/MS (ES+) 592 (M+1) 1 peak at 4.32 min.

[0172] Analytical HPLC(A) t=14.69 min.

[0173] 1H:d-6 DMSO: 0.77 (6H,d), 1.82 (2H,m), 2.06 (1H,m), 2.57 (2H,m),2.82 (4H,m), 2.97 (1H,m), 3.30 (5H,m), 3.55 (1H,m), 3.65 (1H,m), 3.70(1H,m), 3.95 (1H,m), 4.88 (1H,m), 4.95 (1H,m), 6.62 (2H,d), 7.20 (6H,m),7.39 (3H,m), 7.78 (3H,bs).

[0174] 13C (dmso): 155.9, 152.9, 138.5, 129.2, 128.9, 128.1, 126.1,122.9, 112.7, 74.7, 74.5, 72.6, 66.2, 57.2, 53.2, 49.4, 38.8, 37.94,35.1, 32.1, 26.3, 20.0, 19.8.

EXAMPLE 7

[0175]

[0176] The synthesis of compound 204 from compound 198 was carried asdescribed in Example 1, except that mono-1,3-diamino-3-N-Boc-propane wasused in place of di-p-nitrophenyl carbonate.

[0177] C,H,N: calc: 49.07, 5.64, 8.89, found 48.95, 6.00, 8.92C₂₉H₄₃N₅O₇S₁×1.6 CF₃COOH

[0178] LC/MS (ES+) 605 (M+1) 1 peak at 4.27 min.

[0179] Analytical HPLC(A) t=14.72 min.

[0180] 1H:d-6 DMSO: 0.78 (6H,dd), 1.64 (2H,m), 1.83 (2H,m), 2.03 (1H,m),2.57 (1H,m), 2,78 (4H,m), 2.94 (1H,m), 3.03 (2H,m), 3.32 (2H,m), 3.58(1H,m), 3.63 (1H,m), 3.73 (1H,m), 3.87 (1H,m), 4.84 (1H,m), 4.92 (1H,m),6.61 (2H,d), 7.22 (6H.m), 7.36 (1H,d), 7.28 (2H,d), 7.76 (3H,ns).

[0181] 13C (dmso): 155.8, 155.7, 138.5, +129.1, +129.0, +128.0, +126.1,122.9, +112.7, +74.6, +74.3, −72.7, −66.2, −57.2, +53.6, −49.5, −37.4,−36.7, −35.5, −32.1, −27.6, +26.2, +20.0, +19.8.

EXAMPLE 8

[0182]

[0183] The synthesis of compound 205 from compound 198 was carried asdescribed in Example 1, except that 1,4-diamino-4-N-Boc-butane was usedin place of di-p-nitrophenyl carbonate.

[0184] C,H,N: calc: 48.17, 5.59, 8.26, found 48.02, 5.96, 8.24C₃₀H₄₅N₅O₇S₁ 0.2 CF₃COOH

[0185] LC/MS (ES+) 620 (M+1) 1 peak at 4.36 min.

[0186] Analytical HPLC(A)⁻ t=14.93 min.

[0187] 1H: d-6 DMSO: 0.77 (6H,dd), 1.43 (4H,m), 1.82 (2H,m), 2.03(1H,m), 2.77 (4H,m), 2.95 (3H,m), 3.31 (2H,m), 3.56 (1H,m), 3.63 (1H,m),3.70 (1H,bq), 3.82 (1H,m), 4.85 (1H,m), 4.92 (1H,m), 6.62 (2H,d), 7.2(7H,m), 7.38 (2H,d), 7.72 (3H,bs).

[0188] 13C: 155.7, 152.9, +138.6, +129.1, +129.0, +128.0, +126.1,+123.0, +112.7, +74.4, +74.3, −72.7, −66.2, −57.2, +53.7, −49.7, −38.6,−38.5, −35.4, −32.1, −26.3, +26.2, −24.4, +20.1, +19.9.

EXAMPLE 9

[0189]

[0190] The synthesis of compound 206 from compound 198 was carried asdescribed in Example 1, except that (3R)-(+)-3-Boc-aminopyrrolidine wasused in place of di-p-nitrophenyl carbonate.

[0191] C,H,N: calc: 48.28, 5.36, 8.28, found 47.89, 5.53, 8.57C₃₀H₄₃N₅O₇S₁×2 TFA

[0192] LC/MS (ES+) 618 (M+1) 1 peak at 4.32 min.

[0193] Analytical HPLC(A) t=14.31 min.

[0194] 1H and 13C NMR: complex and overlapping mixtures of rotomers.

EXAMPLE 10

[0195]

[0196] The synthesis of compound 207 from compound 198 was carried asdescribed in Example 1, except that (3S)-(−)-3-Boc-aminopyrrolidine wasused in place of di-p-nitrophenyl carbonate,

[0197] LC/MS (ES+) 618 (M+1) 1 peak at 4.19 min.

[0198] Analytical HPLC(A) t=14.75 min.

[0199] 1H and 13C NMR: complex and overlapping mixtures of rotomers.

EXAMPLE 11

[0200]

[0201] The synthesis of compound 308 from compound 198 was carried asdescribed in Example 1, except thatN-triphenylmethyl-N,N′-dimethylethanediamine was used in place ofdi-p-nitrophenyl carbonate.

[0202] 1H-NMR: 0.76 (6H,dd), 1.65 (2H,m), 1.95 (1H,m), 2.07 (1H,m), 2.7(2H,m), 2.75 (3H,s), 2.95 (3H,m), 3.45 (2H,m), 3.7 (4H,m), 4.2 (2H,bm),5.05 (2H,bd), 6.62 (2H,d), 7.2 (5H,m)⁻, 7.5 (2H,d).

[0203] LC/MS: 1 peak, 620 (MH+).

EXAMPLE 12 General Procedures

[0204] Acylation:

[0205] To 200 mg (0.37 mM) of 197 dissolved in 5 ml CH₂Cl₂ was addedN-CBz-L-Benzyl tyrosine 183 mg (0.41 mM) followed by 231 mg (1.12 mM)DCC, followed by 29 mg (0.23 mM) DMAP. The reaction is stirred at rt for24 hr. The precipitates present were removed by filtration. The filtratewas then concentrated in vacuo. The final compound was purified onpreparative reversed phase C18 using purification by HPLC C18 WatersDelta Prep 3000 Column: YMC-Pack ODS AA 12S05-2520WT 250×20 mm I.D. S-5mm, 120 Å, 0-100% B over ½ h, flow=18 ml/min, monitored at 220 nm, B=0.1trifluoroacetic acid in acetonitrile, A=0.1% trifluoroacetic acid inwater. Analytical Column: YMC-Pack ODS AA1 2S05-2520WT 250×4.6 mmI.D.S-5 mm, 120 Å, 0-100% B at 1.5 ml/min. over ½ h, monitored at 220 nm,B=0.1% trifluoroacetic acid in acetonitrile, A=0.1% trifluoroacetic acidin water.

[0206] The aqueous phase was lyophilized to give 59 mg, (16.3%)GW431896X, (U11484-72-10) tHPLC=11.71 min., MW=966.04, LC/MS=MH+967.

[0207] Reduction of the Nitro Functionality:

[0208] A slurry of 209 (170 mg) and 10 mg of 10% Pd.C in 95% EtOH wasflushed with hydrogen in a scintillation vial equipped with septum and astir bar. Continuous overnight hydrogenolysis under hydrogen balloonresulted in a complete conversion. The crude preparation was thenfiltered off the catalyst, and purified on RP C18 HPLC (Prep Nova-PackC186 um, 60 A, gradient 0-100% B over 30 min. The desired product wascollected and lyophilized affording a white fluffy solid (50 mg, 30.8%).

EXAMPLE 13

[0209]

[0210] Compound 211 was obtained following the acylation and reductionprocedures of Example 12. ES+669.2 (M+1), tHPLC=8.06 min (D), 13C NMR(DMSO)168.9, 156.9, 155.7, 153.1, 138.1, 130.5, 129.2, 129.1, 128.1,126.2, 124.7, 122.5, 112.8, 76.2, 74.5, 72.5, 66.1, 58.0, 53.6, 52.6,49.2, 33.6, 32.1, 26.6, 25.3, 20.0. tHPLC=11.71 min (D), ES+ 967 (M+1).

EXAMPLE 14

[0211]

[0212] 212 was obtained following the procedures of Example 12.

[0213] tHPLC=9.45 min (D), ES+592.2 (M+1).

[0214] 13C NMR (DMSO) 171.5, 155.8, 148.9, 137.8, 129.5, 129.3, 128.5,126.7, 115.2, 75.2, 73.8, 73.1, 68.3, 67.0, 58.7, 57.1, 53.3, 49.2,35.4, 32.4, 26.7, 20.1, 19.8.

[0215] 1H(CDCl3, 399.42 KHz): 8.33 (2H, d, J=8.8), 7.95 (2H, d, J=8.8),7.23 (5H, m) 5.22 (m, 2H), 5.08 (m, 1H), 4.08 (m, 1H), 3.80-3.45 (7H,m), 3.41 (3H, s), 2.98 (m, 3H), 2.66 (m, 1H), 2.57 (m, 2H), 2.10 (s,1H), 1.93 (2H, m), 0.82 (3H, d), 0.78 (3H, d).

[0216] ES+ 622 (M+1), 644 (M+Na) tHPLC=10.29 min (D).

[0217] 13C NMR (CDCl3): 171.3, 155.5, 149.9, 145.6, 136.9, 129.2, 128.6,128.5, 126.8, 124.4, 76.7, 75.3, 73.2, 72.9, 68.2, 66.9, 58.7, 55.9,53.1, 48.3, 35.3, 32.7, 26.3, 19.9, 19.8.

EXAMPLE 15

[0218]

[0219] 213 was obtained following the procedure of Example 12.tHPLC=9.21 min (D); ES+ 622 (M+1).

[0220] 13C NMR (CDCl3): 170.54, 156.2, 148.6, 136.8, 129.4, 129.2,128.6, 126.6, 115.7, 76.7, 74.6, 73.2, 71.8, 70.6, 68.2, 66.9, 58.9,57.3, 53.8, 49.4, 36.2, 33.1, 26.8, 19.8, 19.5.

[0221] Intermediate: t HPLC=10.05 min (D); ES+=652 (M+H) 674 (M+Na).

EXAMPLE 16

[0222]

[0223] 214 was obtained following the procedure of Example 12.

[0224] ES+ 634.4 (M+1); t HPLC=7.17 min (D).

[0225] 13C (DMSO): 169.3, 155.8, 153.1, 138.0, 129.1, 129.0, 128.1,126.3, 122.6, 112.8, 94.3, 75.6, 74.6, 72.4, 66.1, 57.8, 52.7, 52.0,49.3, 38.4, 34.7, 32.2, 29.1, 26.6, 21.4, 20.1, 20.0.

EXAMPLE 17

[0226]

[0227] 215 was obtained following the procedure of Example 12.

[0228] t HPLC=9.12 min (D)

[0229] 1H (DMSO) all signals broad: 7.38 (3H, br m), 7.20 (5H, br m) ,6.62 (2H, br m), 5.15 (1H, br m), 4.92 (1H, br m), 4.00 (3H, m), 3.7-3.0(16H, m), 2.78 (2H, m), 2.57 (3H, m), 2.04 (m, 1H), 1.78 (m, 2H), 0.77(6H, m)

[0230] 13C (DMSO) 170.6, 156.3, 153.7, 139.1, 129.8, 128.4, 126.7,123.7, 113.3, 79.8, 79.2, 77.3, 76.1, 75.4, 75.2, 73.0, 71.9, 52.3,51.8, 48.2, 46.7, 39.9, 38.7, 25.8, 22.6.

[0231] Intermediate:

[0232] t HPLC=10.18 min (D); ES+ 696.3 (M+1).

EXAMPLE 18

[0233]

[0234] 216 was obtained following the procedure of Example 12.

[0235] 1H-NMR: 0.97 (6H,t), 1.95 (2H,m), 2.20 (1H,m), 2.9 (2H,m), 2.96(6H,s), 3.00 (3H,s), 3.38 (1H,m), 3.42 (3H,m), 3.36 (1H,m), 3.6 (2H,m),3.7 (6H,m), 3.98 (2H,m), 4.2 (2H,dd), 5.1 (1H,bs), 5.4 (1H,m), 6.8(2H,d), 7.4 (5H,m), 7.6 (2H,d).

[0236] LC-MS: 1 peak, 692 (MH+).

EXAMPLE 19

[0237]

[0238] 217 was obtained following the procedure of Example 12.

[0239] 1H-NMR (CDCl3): 0.78 (6H,dd), 1.9 (2H,m), 2.1 (1H,m), 2.3 (3H,s),2.9 (8H,m), 2.9 (2H,m), 3.15 (1H,m), 3.35 (1H,m), 3.5 (1H,m), 3.75(4H,m), 4.06 (2H,s), 4.15 (2H,m), 4.9 (1H,dd), 5.05 (1H,bs), 5.2(1H,bs), 6.63 (2H,d), 7.2 (5H,m), 7.55 (2H,d), 8.0 (2H,m).

[0240] ESMSP: 676 (MH+).

EXAMPLE 20 General Procedure for N-acylated Compounds

[0241]

[0242] A mixture of 0.5 g (1 mMol) of(3S)-Tetrahydro-3-furfuryl-N-((1S,2R)-1-benzyl-2-hydroxy-3-(N-isobutyl-4-aminobenzenesulfonamido)propyl)carbamate, 0.4 g (1.5 mMol) of Boc-(S)-3-pyridyl alanine, 0.29 g (1.5mMol) EDCI and 0.1 g 4-dimethylamino pyridine in 10 ml ofN,N-dimethylformamide was stirred at 25° for 12 hours. The volatileswere removed in vacuo and the residue was partitioned between ethylacetate and 1N hydrochloric acid. The organic layer was washed with 1Nsodium hydroxide and brine, dried over magnesium sulfate andconcentrated in vacuo. The residue was chromatographed on a 2 inch plugof silica gel (1:1 ethyl acetate: hexane) to give the desired N-acylatedmaterial. Deprotection by treatment with 50 ml of trifluoroacetic acid,followed by co-evaporation of residual acid with methanol gave thedesired prodrug as a white foam (0.2 g, 26%).

[0243] H1-NMR (acetonitrile-D3): 0.95 (6H,dd), 2.0 (2H,m), 2.25 (1h,m),2.8-3.1 (5H,m), 3.6-4.0 (7H,m), 4.25 (1H,m), 4.75 (1H,m), 5.18 (1H,m),5.45 (1H,m), 7.0 (2H,d), 7.4 (5H,m), 7.75 (2H,d), 8.2 (1H,m), 8.8(1H,d), 8.85 (1H,d), 9.15 (1H,s).

[0244] LC/MS: 1 peak, 654 (MH+).

EXAMPLE 21

[0245]

[0246] 220 was obtained using the general procedure in Example 20.

[0247] 1H-NMR (acetone-d6/methanol-d4): 0.95 (6H,t), 2.0 (2H,m), 2.2(1H,m), 2.90 (1H,dd), 2.95 (2H,d), 3.12 (1H,dd), 3.4 (2H,m), 6 (1H,d),3.8 (5H,m), 4.4 (2H,bm), 6.82 (2H,d), 7.20 (1H,s), 7.4 (5H,m), 7.65(2H,d), 8.0 (1H,s).

[0248] LC/MS: 1 peak, 643 (MH+).

EXAMPLE 22

[0249]

[0250] 221 was obtained using the general procedure in Example 20.

[0251] 1H-NMR (DMSO d-6): 0.76 (6H,t), 1.80 (2H,m), 2.10 (1H,m), 3.7(4H,m), 3.75 (3H,s), 3.2 (5H,m), 3.58 (2H,s), 3.7 (4H,m), 4.97 (1H,bm),5.18 (1H,bs), 6.7 (2H,d), 7.22 (5H,m), 7,45 (2H,d).

[0252] LC/MS: 1 peak, 646 (MH+).

EXAMPLE 23

[0253]

[0254] 222 was obtained using the general procedure in Example 20.

[0255] 1HNMR (acetonitrile d-3): 1.0 (6H,t), 2.0 (2H,m), 2.2 (1H,m),3.00 (6H,s), 3.02 (3H,s), 3.1 (4H,m), 3.5 (3H,m), 3.8 (8H,m), 4.4(2H,s), 5.15 (1H,bs), 7.4 (5H,m), 7.97 (2H, d), 8.04 (2H,d),

[0256] LC/MS: 1 peak, 692 (MH+).

EXAMPLE 24

[0257]

[0258] 223 was obtained using the general procedure in Example 20.

[0259] t HPLC=9.22 min (D); ES+ 622 (M+1).

[0260] 1H NMR d6-DMSO: 0.76 (6H,dd), 1.0-1.8 (15H,m), 2.03 (1H,m), 2.58(2H,m), 2.79 (2H,m), 3.11 (1H,m), 3.28 (3H,s), 3.3-3.5 (12H,m), 3.94(1H,m), 4.08 (1H,m), 4.94 (1H,m), 5.14 (1H,m), 6.61 (2H,d), 7.22 (5H,m),7.40 (3H,m).

[0261] 13C (DMSO)169.7, 165.9, 152.9, 138.4, 129.2, 129.1, 128.1, 126.2,123.1, 112.8, 74.4, 74.1, 72.5, 71.2, 69.8, 66.1, 58.1, 57.1, 52.9,47.5, 33.4, 33.2, 26.3, 24.5, 18.9, 18.8.

EXAMPLE 25

[0262]

[0263] 224 was obtained using the general procedure in Example 20.

EXAMPLE 26 O, N-diacylated Prodrugs

[0264] The general procedure for N,O-diacylated compounds followed theprotocol outlined in Example 20, above, except that a five fold excessof reagents was used relative to the starting material.

[0265] t HPLC 9.26 min (D); ES+ 738 (M+1) 760 (M+Na).

[0266] 13C (DMSO): 170.2, 169.8, 156.4, 143.4, 138.8, 129.5, 128.8,128.5, 126.8, 119.7, 74.9, 74.2, 73.7, 71.6, 70.7, 70.3, 68.0, 67.2,59.3, 57.6, 53.8, 49.6, 35.7, 33.8, 27.1, 20.4.

[0267] 1H (DMSO): 10.1 (1H, s), 7.84 (d, 2H, J=8.5), 7.76 (d, J=8.7,2H), 7.40 (1H, d, J=9.2), 7.22 (m, 5H), 5.14 (1H, m), 4.95 (1H, m), 4.1(m, 8H), 3.7-3.3 (m, 13H), 3.28 (s, 3H), 3.26 (s, 3H), 2.86 (m, 2H),2.73 (m, 1H), 2.59 (m, 1H), 2.04 (m, 1H), 1.83 (m, 2H), 0.78 (m, 6H).

EXAMPLE 27

[0268]

[0269] To a mixture of 197 (2.93 g, 5.47 mmol) and phosphorous acid(Aldrich, 2.2 equiv., 12.03 mmol, 987 mg) in 20 ml pyridine was added1,3-dicyclohexylcarbodiimide (Aldrich, 2.1 equiv., 11.49 mmol, 2.37 g)and the reaction heated to 60° C. under nitrogen for 3 h. Solvent wasremoved in vacuo, the residue treated with 200 ml 0.1N aqueous sodiumbicarbonate and stirred 1 h at ambient temperature. The mixture wasfiltered, the filtrate acidified to pH 1.5 by addition of conc. HCl andextracted with ethyl acetate (3×100 ml). The combined orgnic layers weredried over magnesium sulfate, filtered and concentrated in vacuo to give3.15 g (96%) of desired product 226 which was used directly in the nextreaction. HPLC: Rt=8.91 min (96%), MS (AP+) 600.5 (M+1).

EXAMPLE 28

[0270]

[0271] A suspension of 226 (˜5.47 mmol) in 18 ml hexamethyldisilazanewas stirred at 120° C. until homogeneous followed by addition ofbis(trimethylsilyl) peroxide (Gelest, Inc., 2.3 equiv., 12.58 mmol, 2.24g, 2.71 ml). After 1 h the mixture was cooled to ambient temperature,solvent removed in vacuo, the residue stirred with 100 ml methanol,solvent removed in vacuo, the residue stirred with 100 ml 0.1N aqueoussodium bicarbonate, acidified to pH 1.5 by addition of conc. HCl,saturated with brine and extracted with ethyl acetate (3×100 ml). Thecombined organic layers were dried over magnesium sulfate, filtered andconcentrated in vacuo to give 2.98 g (88%) of desired product 227, whichwas used directly in the next reaction. HPLC: Rt=9.28 min (90%), MS(AP+) 616.5 (M+1).

[0272] Alternatively, 227 can be synthesized directly from 197. In thismethod, 197 was dissolved in pyridine (300 mL). The resulting solutionwas concentrated in vacuo to about 150 ml at 50-55° C. The solution wasthen cooled under N₂ to 5° C., and treated with POCl₃ (6.5 ml, 1.24equiv.) over 2 minutes. The cooling bath was removed and the reactionstirred at ambient temperature for 2.5 hrs. The solution was then cooledto 5° C. and water (300 ml) was added over 30 minutes.

[0273] The resulting mixture was extracted with 4-methylpentan-2-one(MIBK, 2×150 ml). The combined extracts were washed with 2N HCl (2×250ml). The acid washes were back extracted with MIBK (60 ml), then thecombined MIBK solutions were treated with 2N HCl (150 ml). The two phasemixture was stirred rapidly and heated to 50° C. for 2 hours. Thereaction mixture was cooled to 20° C., the phases were separated and theMIBK solution was washed with brine (150 ml). The product, 227, wasisolated by drying the solution with magnesium sulfate, filtering of thedrying agent and concentrating in vacuo at 40° C. to give the product asa pale yellow foam (31 g, 90% yield)

EXAMPLE 29

[0274]

[0275] A solution of 227 (2.98 g, 4.84 mmol) in 50 ml ethyl acetate wastreated with 10% palladium on carbon (Aldrich, 300 mg) and put under 35psi of hydrogen on a Parr shaker for 15 h. Catalyst was removed byfiltration and solvent removed in vacuo to give 2.66 g (94%) of desiredproduct 228. HPLC: Rt=7.23 min (92%), MS (ES+) 586.3 (M+1).

EXAMPLE 30

[0276]

[0277] Solid 228 (2.66 g, 4.54 mmol) was treated with 10 ml aqueoussodium bicarbonate (Baker, 3.0 equiv., 13.63 mmol, 1.14 g) and loadedonto a resin column (Mitsubishi Kasei Corp., MCI-gel, CHP-20). Distilledwater was run through until the eluent was neutral followed by productelution with 1 acetonitrile in water. Pure fractions were pooled andlyophilized to give 918 mg of pure bis-sodium salt 229.

EXAMPLE 31

[0278]

[0279] 0.53 g (3.0 mmol) 2-[2-(2-Methoxyethoxy)ethoxy] acetic acid wasadded to a stirred solution of 1.2 g (3.15 mmol) HATU 0.2 g (1.47 mmol)HOAt 0.4 g (4.0 mmol) NMM in 10 ml anhydrous N,N-dimethylformamide. Themixture was stirred at room temperature for 30 minutes, then 0.5 g (1mmol) of(3S)-Tetrahydro-3-furfuryl-N-((1S,2R)-1-benzyl-2hydroxy-3-(N-isobutyl-4-aminobenzenesulfonamido)-propyl)carbamate was added to the solution in one portion. The mixture wasstirred at 20° C. for an hour then at 50° C. for an additional 12 hours.It was then cooled to 20° C., 50 ml of ether was added, and the solutionwas washed with water three times. The aqueous phase was washed withether, and then the combined organic phases were dried with anhydrousmagnesium sulfate and filtered. The filtrate was concentrated underreduced pressure and the residue was purified by silica gelchromatography to obtain the desired Mono-(N)acylated (102 mg, 15%) andBis-(O,N) acylated (262 mg, 32%) compounds.

[0280] Mono-(N)-acylated: 1H-NMR(CDCl3): 0.85 (dd, 6H), 1.85 (m, 2H),2.08 (m,1H), 2.8-3.1 (m, 7H), 3.33 (s, 3H), 3.55 (m, 3H), 3.70-3.90 (m,8H), 4.1 (s, 2H), 5.0 (d, 1H), 5.08 (s(br), 1H), 7.2 (m, 5H), 7.70 (d,2H), 7.80 (d, 2H), 9.09 (s, 1H).

[0281] MS(FAB+) : 666 (M+1).

[0282] Bis-(O,N)-acylated: 1H-NMR(CDCl3): 0.77 (m, 6H), 1.81 (m, 1H),1.95 (m, 1H), 2.05 (m, 1H), 2.6-3.0 (m, 6H), 3.2 (m,1H), 3.332 (s, 3H),3.338 (s, 3H), 3.5-3.8 (m, 18H), 4.1 (s, 2H), 4.14 (s, 2H), 4.17 (m,1H), 5.05 (m, 2H), 5.25 (s(br), 1H), 7.2 (m,5H), 7.69 (d, 2H), 7.78 (d2H), 9.06 (s, 1H).

[0283] MS(FAB+): 826(M+1), 848(M+Na).

EXAMPLE 32

[0284]

[0285] We dissolved 0.521 g (1 mM) of 1273W94 in 5 ml THF, then cooledto −78° C. under nitrogen, and added 1.56 ml (2.5 mM) of a 1.6 Msolution of nBuLi in hexane. After 20 min at −78° C., we added 105 μL(1.1 mM) of ethyl chlorocarbamate and warmed up the reaction to roomtemperature, followed by addition of another 105 μL of ethylchlorocarbamate.

[0286] After stirring for additional 4 hrs, the reaction was quenchedwith water and the organic solvent evaporated. Part of the crude productwas purified on a silica gel (Rf=0.69 (1:2 ethyl acetate:hexane)),yielding 0.131 g of the product.

[0287] C,H,N: calc: 46.06, 4.97, 5.88, found 45.90, 4.97, 5.88C₂₃H₃₃N₅O₅S₁. 2.2 TFA

[0288] LC/MS (ES+) 594 (M+1) 1 peak at 6.96 min.

[0289] Analytical HPLC(A) t=24.57 min.

[0290] 13C (CDCl3): 155.8, 154.4, 149.9, 145.7, 136.8, +129.2, +128.7,+126.8, +124.2, 80.1, +76.9, −64.3, −56.2, −52.5, −48.7, −36.2, +28.1,+26.4, +20.0, +19.8, +14.3.

EXAMPLE 33

[0291]

[0292] We dissolved 0.131 g of the above ethyl carbonate in 4 ml DCM,followed by 4 ml of TFA. Solvents were then removed after 45 min at roomtemperature, resulting in the title compound.

[0293] 1H (DMSO): 8.37 (2H, d, J=7.2), 8.15 (2H, m), 8.00 (2H, d,J=7.0), 7.37 (5H, m), 5.04 (1H, d, J=6.9), 4.06 (2H, q, J=7.0), 3.82((1H, m), 3.35 (2H, m), 2.95 (4H, m), 1.82 (1H, m), 1.20 (3H, t, J=7.0),0.72 (overlapping doublets, 6H, J=6.2).

[0294] LC/MS 1 peak at 4.76 min.

[0295] ES+ 497.3 (M+1).

EXAMPLE 34 O, N-Acyloxy Rearrangement

[0296]

[0297] C,H,N: calc:53.26, 6.14, 7.57, found 53.22, 6.14, 7.57C₂₃H₃₃N₅O₅S₁×0.8 TFA

[0298] LC/MS (ES+) 594 (M+1) 1 peak at 6.96 min.

[0299] Analytical HPLC(A) t=24.57 min.

[0300] 1H (DMSO): 8.34 (2H, d, J=8.7), 8.02 (2H, d, J=8.0), 7.19 (5H,m), 6.98 (1H, d, J=7.2), 5.00 (1H, m), 3.83 (2H, q), 3.50 (2H, m), 3.06(m, 2H), 2.96 (2H, m), 2.43 (1H, m), 1.97 (1H, m), 1.02 (3H, t), 0.84(3H, d), 0.82 (3H, d). 13C (DMSO): 156.2, 150.1, 145.7, 140.0, +129.7,+129.2, +128.5, +126.3, +125.0, +71.8, −60.0, +56.2, −56.0, −51.8,−36.0, +26.3, +20.3, +20.1, +14.6.

EXAMPLE 35

[0301]

[0302] Synthesis of 235 was accomplished analogous to that set forth inExample 1.

[0303] Yield 15.2%; tHPLC=25.2 min (A).

[0304] Rf=0.54 (B); ES+ 687.3 (M+1).

[0305] 1H (CDCl3): 8.34 (overlapping d+d, 4H), 7.97 (d, 2H, J=8.9), 7.35(7H, m), 5.09 (1H, m), 4.56 (1H, d, J=8.4), 4.20 (1H, m), 3.54 (1H, m),3.00 (3H, m), 2.82 (1H, m), 1,84 (1H, m), 1.37 (9H, s), 0.84 (3H, d),0.82 (3H, d).

EXAMPLE 36

[0306]

[0307] We dissolved 150 mg of 235 in 3 ml of anhydrous dioxane, added0.35 ml of S(+)-3-OH-THF and 0.14 ml triethyl amine. The mixture wasrefluxed gently under nitrogen for 2 days. Conversion to 236 wasquantitative. Solvents were removed and the compound purified on silica(B).

[0308] tHPLC=22.98 min (A); ES+ 636.2 (M+1).

[0309] 1H NMR (CDCl3): 8.29 (2H, d), 7.91 (2H, d), 7.22 (5H, m), 5.13(1H, m), 4.96 (1H, m), 4.52 (1H, d), 4.02 (1H, m), 3.84 (2H, m), 3.44(1H, m), 3.36 (1H, m), 3.10 (3H, m, overlap), 2.88 (2H, m), 2.64 (1H,m), 2.14 (1H, m), 2.05 (1H, m), 1.84 (1H, m), 1.27 (9H, s), 0.78 (6H,two overl. d).

EXAMPLE 37 Carbohydrate-Based Prodrugs

[0310]

[0311] A mixture of 0.54 g (1 mMol) of(3S)-Tetrahydro-3-furfuryl-N-((1S,2R)-1-benzyl-2-hydroxy-3-(N-isobutyl-4-aminobenzenesulfonamido)propyl)carbamate, 0.46 g (2 mMol) of 5-dimethyl-tert-butyosilyloxypentanoicacid, 0.346 g (1.8 mMol) of EDCI and 0.556 mL (4 mMol) of triethylaminein 10 ml of dimethyl formamide was stirred at rt for 24 h. Another 3mMol each of acid, EDCI and triethylamine were added and stirring wascontinued for an additional 96 h. A third batch of acid and EDCI wasadded (3 mMol each) and the mixture was stirred 72 h to complete thereaction.

[0312] The reaction mixture was then diluted with ethyl acetate andextracted with 1 N hydrochloric acid, saturated sodium bicarbonate andwater. Evaporation of the solvent and purification on silica gel (30%ethyl acetate-hexane) gave the desired product (500 mg) as a waxy solid.

[0313] LCMS: 1 peak, 772.5 (M+Na)

[0314] 1H NMR (CDCL3): 0.01 (6H,s), 0.78 (6H,dd), 0.95 (9H,s), 1.4-1.8(6H,m), 1.9 (2H,m), 2.05 (1H,m), 2.3 (2H,m), 2.65 (1H,m), 2.95 (2H,m),3.22 (1H,m), 3.4 (1H,m), 3.6 (2H,m), 3.75 (3H,m), 4.8 (1H,d), 5.1(1H,bs), 5.2 (1H,bs), 7.2 (5H,m), 7.95 (2H,d), 8.36 (2H,d).

[0315] 450 mg of the 238 was dissolved in 30 ml of tetrahydrofuran andtreated with 20 ml of water and 50 ml of acetic acid. The mixture wasstirred at rt for 2 h and evaporated. Titration with hexane gave thedesired alcohol (290 mg) as a white solid.

[0316] A mixture of 0.15 g (0.24 mMol) of the alcohol produced abovefrom the previous reaction, 0.205 g (0.5 mMol) oftetraacetylglucosylbromide and 0.191 g (0.7 mMol) of silver carbonate in3 ml of dichloromethane was stirred at rt for 6 h. 150 mg of additionalglucosyl bromide and 150 mg of silver carbonate were added and themixture was stirred at rt overnight. The mixture was loaded onto a padof silica gel and eluted with 30% ethylacetate-hexane to afford thedesired protected carbohydrate pro-drug as a white foam (200 mg).

[0317] LCMS: 1 peak, 966 (M+H).

[0318] 1H-NMR (CDCl3): 0.78 (6H,dd), 1.9 (2H,m), 2.00 (3H,s), 2.02(3H,s), 2.05 (3H,s), 2.06 (3H,s), 2.1 (2H,m), 2.3 (2H,m), 2.7 (1H,m),2.94 (3H,bd), 3.35 (2H,m), 3.45 (2H.m), 3.8 (5H,m), 4.1 (3H,m), 4.5(1H,d), 4.9 (1H,bs), 4.95 (1H,t,), 5.08 (4H,m), 2H,d), 8.35 (2H,d).

EXAMPLE 38

[0319]

[0320] 1.5 g (9.4 mmol) SO3.py complex was added to a stirred solutionof 1 g (1.87 mmol) of 197 in 25 mL anhydrous tetrahydrofurane. Themixture was stirred at 20° C. for 12 hours, then filtered. The filtratewas concentrated at reduced pressure, and the residue was transferred toa silica gel column and eluted with EtOAc (neat), followed by EtOAc:EtOH(4:1) to obtain 471 mg (47%) 239 as a colorless foam.

[0321] 1H-NMR(CDCl3): 0.80 (m, 6H), 1.8-2.1 (m, 3H), 4.15 (s(br), 1H),4.8 (t, 1H), 5.04 (s (br), 1H).

[0322] MS (ES−): 614 (M−1)

[0323] 100 mg (0.162 mmol) 239 dissolved in 15 ml anhydroustetrahydrofuran and 200 mg Pd/BaSO4 (5%) was added to the solution. Themixture was stirred under atmospheric pressure of hydrogen for 8 hours,and then the catalyst was filtered. The filtrate was concentrated underreduced pressure then dried under vacuum (˜1 Hg mm, 48 hrs.) to produce80 mg (81%) 240 as a colorless foam. 1H-NMR(DMSO-d6): 0.85 (dd, 6H),0.90 (m, 1H), 2.05 (m, 2H), 2.58 (m, 3H), 2.84 (dd, 1H), 3.05 (m, 2H),3.55-3.80 (m, 6H), 4.20 (t, 1H), 4.42 (m, 1H), 4.93 (s(br), 1H), 6.09(s, 2H), 6.70 (d, 2H), 6.80 (d, 1H), 7.15-7.40 (m, 4H), 7.51 (d, 2H).

[0324] MS(ES−): 584 (M−1).

EXAMPLE 39

[0325]

[0326] 780 mg (3 mmol) 2-Chloro-1,3,2-dioxaphospholane was added to astirred solution of 1.07 g (2 mmol) 197 and 0.7 ml (4 mmol)N,N-Diisopropylethylamine in 25 ml dichloromethane at 0° C. The mixturewas allowed to warm up to room temperature and it was stirred for 2hours. The mixture was then cooled to 0° C. and 1.5 g (9.3 mmol) brominewas added in 5 ml dichloromethane. The mixture was stirred for 1 hour at20° C., followed by evaporation under reduced pressure. An aqueoussolution (50%) of 15 ml trimethylamine was added to the residue, and themixture was stirred at 20° C. for 12 hours.

[0327] Solvents were removed under reduced pressure and 50 ml EtOAc:EtOH(9:1) was added to the residue. The solid was filtered, washed withEtOAc:EtOH (9:1) then the filtrate was concentrated under reducedpressure. The residue was chromatographed on a 3 inch plug of silica gelusing ethyl acetate (neat), then methanol (neat), as eluents to obtain1.15 g (82%) 241 as an off-white solid.

[0328] 1H-NMR(CDCl3): 0.60 (dd, 6H), 1.70 (m, 1H), 1.95 (m, 1H), 2.10(m, 1H), 2.8-3.2 (m, 6H), 3.4 (s (br), 9H), 5.09 (s(br), 1H), 7.25 (m,5H), 7.83 (d, 2H), 8.28 (d, 2H). MS(ES+): 701 (M+1), 184 (phosphatidylcholine+).

EXAMPLE 40

[0329]

[0330] 250 mg PdC (10%) was added to a solution of 250 mg (0.35 mmol)241 in 10 ml methanol, and the mixture was stirred under atmosphericpressure of hydrogen for 4 hours at 20° C. The mixture was filtered, andthe filtrate was concentrated under reduced pressure. The residue wasthen dissolved in 10 ml water and lyophilized to obtain 174 mg (74%) 242as white solid.

[0331] 1H-NMR(DMSO-d6): 0.82 (dd, 6H), 1.80-2.00 (m, 2H), 2.10 (m, 1H),2.80 (m, 3H), 3.00 (m, 2H), 3.2 (s (br), 9H), 4.0-4.3 (m, 4H), 4.91(s(br), 1H), 6.08 (s(br), 2H), 6.67(d, 2H), 7.30 (m, 5H), 7.48 (d, 2H),8.12 (d, 1H).

[0332] MS(ES+): 671 (M+1), 184 (phosphatidyl choline+).

EXAMPLE 41

[0333]

[0334] 0.175 ml (2 mmol) phosphorus trichloride was added to a stirredsolution of 1.07 g (2 mmol) 197 and 0.35 ml (2 mmol)N,N-Diisopropylethylamine in 25 ml dichloromethane at 20° C. The mixturewas stirred for 4 hours at 20° C., then 1 ml water was added and stirredfor an additional 12 hours at 20° C. 3 g anhydrous magnesium sulfate wasadded to the mixture and it was stirred for 30 minutes, then filtered.The filtrate was concentrated under reduced pressure and purified bysilica gel chromatography using EtOAc:Hexane (4:1), then EtOAc:EtOH(1:1), to obtain 402 mg (48%) 226 and 427 mg (36%) 243. 226:

[0335] 1H-NMR(DMSO-d6): 0.82 (dd, 6H), 1.84 (m, 1H), 1.98 (m, 1H), 2.10(m, 1H), −2.68 (dd, 1H), 2.9-3.2 (m, 4H), 3.6-3.8 (m, 3H), 3.94 (t, 1H),4.30, (s(br), 1H), 4.97 (s(br), 1H), 7.30 (m, 5H), 8.14 (d, 2H), 8.43(d, 2H).

[0336] MS(ES−): 598 (M−1).

[0337] 243: (1:1 mix of diastereomers):

[0338] 1H-NMR(CDCl3): 0.80 (m, 6H), 1.8-2.1 (m, 4H), 2.8-3.2 (m, 6H),3.7-3.9 (m, 4H), 4.15 (m, 1H), 4.8-5.15 (m, 2H), 5.57, 5.72 ((d,d), 1H),7.25 (m, 5H), 7.95 (dd, 2H), 8.35 (m, 2H).

[0339] MS(ES−): 580 (M−1), 598 ((M+H2O)−1).

EXAMPLE 42

[0340]

[0341] The reduction was carried out as described in Example 40; (Yield:79%).

[0342] 1H-NMR(DMSO-d6): 0.81 (dd, 6H), 1.82 (m, 1H), 1.95 (m, 1H), 2.08(m, 1H), 2.6-3.15 (m, 6H), 3.6-3.75 (m, 3H), 4.03 (t, 1H), 4.28, (m,1H), 4.96 (s(br), 1H), 6.07 (s, 2H), 6.65 (d, 2H), 7.25 (m, 5H), 7.42(d, 2H).

[0343] MS (ES−): 568 (M−1).

EXAMPLE 43

[0344]

[0345] The reduction was carried out as described in Example 40; (Yield;98%).

[0346] (1:1 mix of diastereomers):

[0347] 1H-NMR(DMSO-d6): 0.82 (m, 6H), 1.75-2.0 (m, 2H), 2.05 (m, 1H),2.6-3.2 (m, 6H), 3.55-3.8 (m, 4H), 4.02, 4.22 (m, t, 1H), 4.75 (m, 1H),4.90, 5.01 ((d,d), 1H), 6.12 (s, 1H), 6.68 (d, 2H), 7.30 (m, 5H), 7.49(d, 2H).

[0348] MS(ES−): 550 (M−1), 568 ((M+H20)−1).

EXAMPLE 44 Pharmacokinetics In Sprague-Dawley Rats Following Single OralDose

[0349] In order to study the pharmacokinetics of the prodrugs of thisinvention, we administered single oral doses of a series of prodrugs ofthis invention, as well as VX-478, to male and female Sprague-Dawleyrats. Administration of molar equivalents of a series of prodrugs ofthis invention in a variety of pharmaceutical vehicles was tested.

[0350] Separate groups of male and female Sprague-Dawley rats(3/sex/group) received oral doses of compound 229 by oral gavage, indifferent vehicles at the same dose equivalent (40 mg/kg molarequivalent of VX-478). The different vehicles for compound 229 were: 1)water; 2) 5/4/1; 3) PEG 400; 4) TPGS/PEG 400; and 5) PEG. The vehiclesfor VX-478 were: 1)33% TPGS/PEG400/PEG; and 2) 12.5% TPGS/PEG 400/PEG.

[0351] Blood samples were collected following administration at varioustime intervals and analyzed for the presence of both compound 229 andits metabolite, VX-478, by HPLC and MS methods. The results of thisstudy are tabulated below (Table IV). TABLE IV Compound 229 229 229 229VX-478 VX-478 vehicle H₂O H₂O:PG:EtOH PEG TPGS/PEG 33% 12.5% 5:4:1 400400/PG TPGS/ TPGS/ PEG PEG 400/PG 400/PG number 3 3 3 3 6 ≧3 of ratsMolar 40 PO 40 PO 40 PO 40 PO 41 PO 50 PO equiv. dose/ 478 Dose (mg/Kg)AUC 11.7 ± 10.6 ± 7.4 ± 8.2 ± 29.6 ± 16.2 ± (ug*hr/m 4.8 7.4 1.8 1.6 5.81.8 1) Cmax 7.1 ± 3.3 ± 3.1 ± 3.0 ± 14.0 ± 6.0 ± (μM) 1.7 0.6 0.3 0.72.2 1.0 half 1.7* 3.4* 2.8* 2.8* 2.5 ± 2.2 ± life 0.9 1.0 (hr) Relative39.5^(†) 35.8^(†) 25.0^(†) 27.7^(†) reference reference Avail. 90.2^(††)81.8^(††) 57.1^(††) 63.3^(††) of VX-478

[0352] We performed a similar study on dogs using both a solid capsuleformulation of compound 229 and an ethanolic/methyl cellulose solutionformulation, as compared to a TPGS-containing solution formulation ofVX-478. The results from this study are presented below in Table V.TABLE V Compound 229 229 VX-478 vehicle solid methyl 22% capsulecellulose TPGS/PEG in 5% 400/PG EtOH/water number of dogs 2 2 >2 Molarequiv. dose/ 17 PO 17 PO 17 PO 478 Dose (mg/Kg) AUC 16.7 ± 2.7 14.2 ±3.2 23.5 ± 7.4 (ug*hr/ml) Cmax (μg/ml)  6.1 ± 1.7  6.3 ± 0.3  6.8 ± 1.1Tmax (hr)  2.3 ± 0.6  0.5 ± 0.5  1.0 ± 0.8 Relative Avail. of 71.1 60.4reference VX-478 (%)

[0353] The results demonstrate that oral administration of compound 229as an aqueous solution resulted in improved bioavailability incomparison to the other vehicles studied, Also, following administrationof compound 229, none of that compound was detected in the first timepoint blood sample (or later samples), suggesting first pass metabolismto VX-478. Comparison of the aqueous dose of compound 229 with the twonon-aqueous formulations used for VX-478 indicated equivalence indelivery as illustrated by the range found for the bioavailability.

[0354] While we have described a number of embodiments of thisinvention, it is apparent that our basic constructions may be altered toprovide other embodiments which utilize the products and processes ofthis invention. Therefore, it will be appreciated that the scope of thisinvention is to be defined by the appended claims, rather than by thespecific embodiments which have been presented by way of example.

We claim:
 1. A compound of formula I:

wherein: each R¹ is independently selected from the group consisting ofC(O)—, —S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)—and —NR²—C(O)—C(O)—; each A is independently selected from the groupconsisting of 5-7 membered monocyclic heterocycles containing from 1-3endocyclic heteroatoms, which may be optionally methylated at the pointof attachment, optionally benzofused, optionally attached through aC₁-C₃ alkyl linker and optionally fused with a 5-7 membered monocyclicheterocycle containing from 1-2 endocyclic heteroatoms, and whereinunmethylated THF is expressly excluded; each Ht is independentlyselected from C₃-C₇ cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; or a5-7 membered saturated or unsaturated heterocycle, containing one ormore heteroatoms selected from N, N(R²), O, S and S(O)_(n); wherein saidaryl or said heterocycle is optionally fused to Q; and wherein anymember of said Ht is optionally substituted with one or moresubstituents independently selected from oxo, —OR², SR², —R²,—N(R²)(R²), —R²—OH, —CN, —CO₂R², —C(O)—N(R²)₂, —S(O)₂—N(R²)₂,—N(R²)—C(O)—R², —C(O)—R², —S(O)_(n)—R², —OCF₃, —S(O)_(n)—Q,methylenedioxy, —N(R²)—S(O)₂(R²), halo, —CF₃, —NO₂, Q, —OQ, —OR⁷, —SR⁷,—R⁷, —N(R²)(R⁷) or —N(R⁷)₂; each Q is independently selected from a 3-7membered saturated, partially saturated or unsaturated carbocyclic ringsystem; or a 5-7 membered saturated, partially saturated or unsaturatedheterocyclic ring containing one or more heteroatoms selected from O, N,S, S(O)_(n) or N(R²); wherein Q is optionally substituted with one ormore groups selected from oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)—R²,—R²—OH, —CN, —CO₂R², —C(O)—N(R²)₂, halo or —CF₃; each R² isindependently selected from the group consisting of H and C₁-C₃ alkyloptionally substituted with Q; each x is independently 0 or 1; each R³is independently selected from the group consisting of H, Ht, C₁-C₆alkyl and C₂-C₆ alkenyl wherein any member of said R³, except H, may beoptionally substituted with one or more substituents selected from thegroup consisting of —OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Ht, —CN,—SR², —CO₂R², NR²—C(O)—R²; each n is independently 1 or 2; G, whenpresent, is selected from H, R⁷ or C₁-C₄ alkyl, or, when G is C₁-C₄alkyl, G and R⁷ are bound to one another either directly or through aC₁-C₃ linker to form a heterocyclic ring; or when G is not present(i.e., when x in (G)_(x) is 0), then the nitrogen to which G is attachedis bound directly to the R⁷ group on —OR⁷; each D and D′ isindependently selected from the group consisting of Q; C₁-C₅ alkyl,which may be optionally substituted with one or more groups selectedfrom C₃-C₆ cycloalkyl, —OR², —R³, —O—Q, —S—Q and Q; C₂-C₄ alkenyl, whichmay be optionally substituted with one or more groups selected from thegroup consisting of C₃-C₆ cycloalkyl, —OR², R³, O—Q and Q; C₃-C₆cycloalkyl, which may be optionally substituted with or fused with Q;and C₅-C₆ cycloalkenyl, which may be optionally substituted with orfused with R⁶; each E is independently selected from the groupconsisting of Ht; —O—Ht; Ht—Ht; —O—R³; —NR²R³; C₁-C₆ alkyl, which may beoptionally substituted with one or more groups selected from the groupconsisting of R⁴ and Ht; and C₂-C₆ alkenyl, which may be optionallysubstituted with one or more groups selected from the group consistingof R⁴ and Ht; C₃-C₆ saturated carbocycle, which is optionallysubstituted with one or more groups selected from R⁴ or Ht; or C₅-C₆unsaturated carbocycle, which is optionally substituted with one or moregroups selected from R⁴ or Ht; each R⁴ is independently selected fromthe group consisting of OR², —C(O)—NHR², S(O)₂—NHR², halo, NR²—C(O)—R²and —CN; each R⁵ is independently selected from the group consisting ofH and C₁-C₄ alkyl optionally substituted with aryl; and each R⁶ isindependently selected from the group consisting of aryl, carbocycle andheterocycle, wherein said aryl, carbocycle or heterocycle may beoptionally substituted with one or more groups selected from the groupconsisting of oxo, —OR⁵, —R⁵, N(R⁵)(R⁵), N(R⁵)—C(O)—R⁵, —R⁵—OH, —CN,CO₂R⁵, C(O)—N(R⁵)(R⁵), halo and CF₃; each R⁷ is independently selectedfrom

wherein each M is independently selected from H, Li, Na, K, Mg, Ca, Ba,—N(R²)₄, C₁-C₁₂-alkyl, C₂-C₁₂-alkenyl, —R⁶; wherein 1 to 4 —CH₂ radicalsof the alkyl or alkenyl group, other than the —CH₂ that is bound to Z,is optionally replaced by a heteroatom group selected from O, S, S(O),S(O₂), or N(R²); and wherein any hydrogen in said alkyl, alkenyl or R⁶is optionally replaced with a substituent selected from oxo, —OR², —R²,N(R²)₂, N(R²)₃, R²OH, —CN, —CO₂R², —C(O)—N(R²)₂, S(O)₂—N(R²)₂,N(R²)—C(O)—R₂, C(O)R², —S(O)_(n)—R², OCF₃, —S(O)_(n)—R⁶,N(R²)—S(O)₂(R²), halo, —CF₃, or —NO₂; M′ is H, C₁-C₁₂-alkyl,C₂-C₁₂-alkenyl, —R⁶; wherein 1 to 4 —CH₂ radicals of the alkyl oralkenyl group is optionally replaced by a heteroatom group selected fromO, S, S(O), S(O₂), or N(R²); and wherein any hydrogen in said alkyl,alkenyl or R⁶ is optionally replaced with a substituent selected fromoxo, —OR², —R², —N(R²)₂, N(R²)₃, —R²OH, —CN, —CO₂R², —C(O)—N(R²)₂,—S(O)₂—N(R²)₂, —N(R²)—C(O)—R₂, —C(O)R², —S(O)_(n)—R², —OCF₃,—S(O)_(n)—R⁶, —N(R²)—S(O)₂(R²), halo, —CF₃, or —NO₂; Z is O, S, N(R²)₂,or, when M is absent, H; Y is P or S; X is O or S; and R⁹ is C(R²)₂, Oor N(R²); and wherein when Y is S, Z is not S; and R⁶ is a 5-6 memberedsaturated, partially saturated or unsaturated carbocyclic orheterocyclic ring system, or an 8-10 membered saturated, partiallysaturated or unsaturated bicyclic ring system; wherein any of saidheterocyclic ring systems contains one or more heteroatoms selected fromO, N, S, S(O)_(n) or N(R²); and wherein any of said ring systemsoptionally contains 1 to 4 substituents independently selected from OH,C₁-C₄ alkyl, O—C₁-C₄ alkyl or OC(O)C₁-C₄ alkyl.
 2. The compoundaccording to claim 1, wherein at least one R⁷ is selected from:


3. The compound according to claim 1, wherein D is benzyl.
 4. Thecompound according to claim 3, wherein A is selected from3-(1,5-dioxane)-O—C(O)—, or3-hydroxy-hexahydrofura[2,3]-furanyl-O—C(O)—; D′ is (C₁-C₄)-alkyl whichis optionally substituted with one or more groups selected from thegroup consisting of (C₃-C₆)-cycloalkyl, —OR², —R³, —O—Q and Q; E is(C6-C₁₀)-aryl optionally substituted with one or more substituentsselected from oxo, —OR², SR², —R², —N(R²)₂, —R²—OH, —CN, —C(O)O—R²,—C(O)—N(R²)₂, —S(O)₂—N(R²)₂, —N(R²)—C(O)—R², —C(O)—R², —S(O)_(n)—R²,—OCF₃, —S(O)_(n)—Q, methylenedioxy, —N(R²)—S(O)₂—R₂, halo, —CF₃, —NO₂,Q, —OQ, —OR⁷, —SR⁷, —R⁷, —N(R²)(R⁷) or —N(R⁷)₂; or a 5-memberedheterocyclic ring containing one S and optionally containing N as anadditional heteroatom, wherein said heterocyclic ring is optionallysubstituted with one to two groups independently selected from —CH₃, R⁴,or Ht; and Ht, insofar as it is defined as part of R³, is defined as inclaim 1 except for the exclusion of heterocycles.
 5. The compoundaccording to claim 4 wherein A is 1,3-dioxanyl.
 6. The compoundaccording to claim 5 wherein A is 1,3-dioxan-5-yl.
 7. The compoundaccording to claim 4, wherein: G is hydrogen; D′ is isobutyl; E isphenyl substituted with N(R⁷)₂; each M is independently selected from H,Li, Na, K, Mg, Ca, Ba, C₁-C₄ alkyl or —N(R²)₄; and each M′ is H or C₁-C₄alkyl.
 8. The compound according to claim 3, wherein: E is a 5-memberedheterocyclic ring containing one S and optionally containing N as anadditional heteroatom, wherein said heterocyclic ring is optionallysubstituted with one to two groups independently selected from —CH₃, R⁴,or Ht.
 9. The compound according to claim 3, wherein: E is Htsubstituted with N(R⁷)₂; R⁷ in the —OR⁷ group is —PO(OM)₂ orC(O)CH₂OCH₂CH₂OCH₂CH₂OCH₃ and both R⁷ in the —N(R⁷)₂ substituent of Htare H; or R⁷ in —OR⁷ group shown in formula XXII is C(O)CH₂OCH₂CH₂OCH₃,one R⁷in the —N(R⁷)₂ substituent of Ht is C(O)CH₂OCH₂CH₂OCH₃ and theother R⁷ in the —N(R⁷)₂ substituent of Ht is H; and wherein M is H, Li,Na, K or C₁-C₄ alkyl.
 10. The compound according to claim 3, wherein R⁷in the —OR⁷ group is —PO(OM)₂ or —C(O)—M′ and M is Na or K.
 11. Thecompound according to claim 2, wherein: R³ is (C₁-C₆)-alkyl,(C₂-C₆)-alkenyl, (C₅-C₆)-cycloalkyl, (C₅-C₆)-cycloalkenyl, or a 5-6membered saturated or unsaturated heterocycle; wherein any member of R³is optionally substituted with one or more substituents selected fromthe group consisting of —OR², —C(O)—NH—R², —S(O)_(n)N(R²)₂, —Ht, —CN,—SR², —C(O)O—R² and N(R²)—C(O)—R²; and D′ is (C₁-C₃)-alkyl or C₃alkenyl; wherein D′ is optionally substituted with one or more groupsselected from (C₃-C₆)-cycloalkyl, —OR², —O—Q or Q.
 12. The compoundaccording to claim 11, wherein R⁷ in the —OR⁷ group is —PO(OM)₂ or—C(O)—M′.
 13. A pharmaceutical composition, comprising a compoundaccording to any one of claims 1 to 12 in an amount effective to treatinfection by a virus that is characterized by an aspartyl protease; anda pharmaceutically acceptable carrier, adjuvant or vehicle.
 14. Thepharmaceutical composition according to claim 13, wherein said virus isHIV.
 15. The pharmaceutical composition according to claim 13, whereinsaid pharmaceutical composition is formulated for oral administration.16. The pharmaceutical composition according to claim 13, furthercomprising one or more agents selected from an anti-viral agent, an HIVprotease inhibitor other than a compound according to claim 1, and animmunostimulator.
 17. The pharmaceutical composition according to claim16, further comprising one or more agents selected from zidovudine(AZT), zalcitabine (ddC), didanosine (ddI), stavudine (d4T), 3TC,935U83, 1592U89, 524W91, saquinavir (Ro 31-8959), L-735,524, SC-52151,ABT 538 (A80538), AG 1341, XM 412, XM 450, CPG 53,437, or tuscarasol.18. A method for inhibiting aspartyl protease activity in a mammal,comprising the step of contacting administering to said mammal apharmaceutical composition according to claim
 13. 19. A method fortreating HIV infection in a mammal comprising the step of administeringto said mammal a pharmaceutical composition according to claim
 13. 20.The method according to claim 19, wherein said mammal is additionallyadministered one or more additional agents selected from an anti-viralagent, an HIV protease inhibitor other than a compound according toclaim 1, and an immunostimulator either as a part of a single dosageform with said pharmaceutical composition or as a separate dosage form.21. The method according to claim 20, wherein said additional agent isselected from zidovudine (AZT), zalcitabine (ddC), didanosine (ddI),stavudine (d4T), 3TC, 935U83, 1592U89, 524W91, saquinavir (Ro 31-8959),L-735,524, SC-52151, ABT 538 (A80538), AG 1341, XM 412, XM 450, CPG53,437, or tuscarasol.
 22. The method according to claim 19, whereinsaid step of administering comprises oral administration.